Transporters and ion channels

ABSTRACT

The invention provides human transporters and ion channels (TRICH) and polynucleotides which identify and encode TRICH. The invention also provides expression vectors, host cells, antibodies, agonists, and antagonists. The invention also provides methods for diagnosing, treating, or preventing disorders associated with aberrant expression of TRICH.

TECHNICAL FIELD

[0001] This invention relates to nucleic acid and amino acid sequencesof transporters and ion channels and to the use of these sequences inthe diagnosis, treatment, and prevention of transport, neurological,muscle, and immunological disorders, and in the assessment of theeffects of exogenous compounds on the expression of nucleic acid andamino acid sequences of transporters and ion channels.

BACKGROUND OF THE INVENTION

[0002] Eukaryotic cells are surrounded and subdivided into functionallydistinct organelles by hydrophobic lipid bilayer membranes which arehighly impermeable to most polar molecules. Cells and organelles requiretransport proteins to import and export essential nutrients and metalions including K⁺, NH₄ ⁺, P_(i), SO₄ ²⁻, sugars, and vitamins, as wellas various metabolic waste products. Transport proteins also play rolesin antibiotic resistance, toxin secretion, ion balance, synapticneurotransmission, kidney function, intestinal absorption, tumor growth,and other diverse cell functions (Griffith, J. and C. Sansom (1998) TheTransporter Facts Book, Academic Press, San Diego Calif., pp. 3-29).Transport can occur by a passive concentration-dependent mechanism, orcan be linked to an energy source such as ATP hydrolysis or an iongradient Proteins that function in transport include carrier proteins,which bind to a specific solute and undergo a conformational change thattranslocates the bound solute across the membrane, and channel proteins,which form hydrophilic pores that allow specific solutes to diffusethrough the membrane down an electrochemical solute gradient.

[0003] Carrier proteins which transport a single solute from one side ofthe membrane to the other are called uniporters. In contrast, coupledtransporters link the transfer of one solute with simultaneous orsequential transfer of a second solute, either in the same direction(symport) or in the opposite direction (antiport). For example,intestinal and kidney epithelium contains a variety of symporter systemsdriven by the sodium gradient that exists across the plasma membrane.Sodium moves into the cell down its electrochemical gradient and bringsthe solute into the cell with it. The sodium gradient that provides thedriving force for solute uptake is maintained by the ubiquitous Na⁺/K⁺ATPase system. Sodium-coupled transporters include the mammalian glucosetransporter (SGLT1), iodide transporter (NIS), and multivitamintransporter (SMVT). All three transporters have twelve putativetransmembrane segments, extracellular glycosylation sites, andcytoplasmically-oriented N- and C-termini. NIS plays a crucial role inthe evaluation, diagnosis, and treatment of various thyroid pathologiesbecause it is the molecular basis for radioiodide thyroid-imagingtechniques and for specific targeting of radioisotopes to the thyroidgland (Levy, O. et al. (1997) Proc. Natl. Acad. Sci. USA 94:5568-5573).SMVT is expressed in the intestinal mucosa, kidney, and placenta, and isimplicated in the transport of the water-soluble vitamins, e.g., biotinand pantothenate (Prasad, P. D. et al. (1998) J. Biol. Chem.273:7501-7506).

[0004] One of the largest families of transporters is the majorfacilitator superfamily (MFS), also called theuniporter-symporter-antiporter family. MFS transporters are singlepolypeptide carriers that transport small solutes in response to iongradients. Members of the MFS are found in all classes of livingorganisms, and include transporters for sugars, oligosaccharides,phosphates, nitrates, nucleosides, monocarboxylates, and drugs. MFStransporters found in eukaryotes all have a structure comprising 12transmembrane segments (Pao, S. S. et al. (1998) Microbiol. Molec. Biol.Rev. 62:1-34). The largest family of MFS transporters is the sugartransporter family, which includes the seven glucose transporters(GLUT1-GLUT7) found in humans that are required for the transport ofglucose and other hexose sugars. These glucose transport proteins haveunique tissue distributions: and physiological functions. GLUT1 providesmany cell types with their basal glucose requirements and transportsglucose across epithelial and endothelial barrier tissues; GLUT2facilitates-glucose uptake or efflux from the liver; GLUT3 regulatesglucose supply to neurons; GLUT4 is responsible for insulin-regulatedglucose disposal; and GLUT5 regulates fructose uptake into skeletalmuscle. Defects in glucose transporters are involved in a recentlyidentified neurological syndrome causing infantile seizures anddevelopmental delay, as well as glycogen storage disease, Fanconi-Bickelsyndrome, and non-insulin-dependent diabetes mellitus (Mueckler, M.(1994) Eur. J. Biochem. 219:713-725; Longo, N. and L. J. Elsas (1998)Adv. Pediatr. 45:293-313).

[0005] Monocarboxylate anion transporters are proton-coupled symporterswith a broad substrate specificity that includes L-lactate, pyruvate,and the ketone bodies acetate, acetoacetate, and beta-hydroxybutyrate.At least seven isoforms have been identified to date. The isoforms arepredicted to have twelve transmembrane (TM) helical domains with a largeintracellular loop between TM6 and TM7, and play a critical role inmaintaining intracellular pH by removing the protons that are producedstoichiometrically with lactate during glycolysis. The bestcharacterized H⁺-monocarboxylate transporter is that of the erythrocytemembrane, which transports L-lactate and a wide range of other aliphaticmonocarboxylates. Other cells possess H⁺-linked monocarboxylatetransporters with differing substrate and inhibitor selectivities. Inparticular, cardiac muscle and tumor cells have transporters that differin their K_(m) values for certain substrates, includingstereoselectivity for L- over D-lactate, and in their sensitivity toinhibitors. There are Na⁺-monocarboxylate cotransporters on the luminalsurface of intestinal and kidney epithelia, which allow the uptake oflactate, pyruvate, and ketone bodies in these tissues. In addition,there are specific and selective transporters for organic cations andorganic anions in organs including the kidney, intestine and liver.Organic anion transporters are selective for hydrophobic, chargedmolecules with electron-attracting side groups. Organic cationtransporters, such as the ammonium transporter, mediate the secretion ofa variety of drugs and endogenous metabolites, and contribute to themaintenance of intercellular pH (Poole, R. C. and A. P. Halestrap (1993)Am. J. Physiol. 264:C761-C782; Price, N. T. et al. (1998) Biochem. J.329:321-328; and Martinelle, K and I. Haggstrom (1993) J. Biotechnol.30:339-350).

[0006] ATP-binding cassette (ABC) transporters are members of asuperfamily of membrane proteins that transport substances ranging fromsmall molecules such as ions, sugars, amino acids, peptides, andphospholipids, to lipopeptides, large proteins, and complex hydrophobicdrugs. ABC transporters consist of four modules: two nucleotide-bindingdomains (NBD), which hydrolyze ATP to supply the energy required fortransport, and two membrane-spanning domains (MSD), each containing sixputative transmembrane segments. These four modules may be encoded by asingle gene, as is the case for the cystic fibrosis transmembraneregulator (CFTR), or by separate genes. When encoded by separate genes,each gene product contains a single NBD and MSD. These “half-molecules”form homo- and heterodimers, such as Tap1 and Tap2, the endoplasmicreticulum-based major histocompatibility (MHC) peptide transport system.Several genetic diseases are attributed to defects in ABC transporters,such as the following diseases and their corresponding proteins: cysticfibrosis (CFTR, an ion channel), adrenoleukodystrophy(adrenoleukodystrophy protein, ALDP), Zellweger syndrome (peroxisomalmembrane protein-70, PMP70), and hyperinsulinemic hypoglycemia(sulfonylurea receptor, SUR). Overexpression of the multidrug resistance(MDR) protein, another ABC transporter, in human cancer cells makes thecells resistant to a variety of cytotoxic drugs used in chemotherapy(Taglicht, D. and S. Michaelis (1998) Meth. Enzymol. 292:130-162).

[0007] A number of metal ions such as iron, zinc, copper, cobalt,manganese, molybdenum, selenium, nickel, and chromium are important ascofactors for a number of enzymes. For example, copper is involved inhemoglobin synthesis, connective tissue metabolism, and bonedevelopment, by acting as a cofactor in oxidoreductases such assuperoxide dismutase, ferroxidase (ceruloplasmin), and lysyl oxidase.Copper and other metal ions must be provided in the diet, and areabsorbed by transporters in the gastrointestinal tract. Plasma proteinstransport the metal ions to the liver and other target organs, wherespecific transporters move the ions into cells and cellular organellesas needed. Imbalances in metal ion metabolism have been associated witha number of disease states (Danks, D. M. (1986) J. Med. Genet.23:99-106).

[0008] Transport of fatty acids across the plasma membrane can occur bydiffusion, a high capacity, low affinity process. However, under normalphysiological conditions a significant fraction of fatty acid transportappears to occur via a high affinity, low capacity protein-mediatedtransport process. Fatty acid transport protein (FATP), an integralmembrane protein with four transmembrane segments, is expressed intissues exhibiting high levels of plasma membrane fatty acid flux, suchas muscle, heart, and adipose. Expression of FATP is upregulated in3T3-L1 cells during adipose conversion, and expression in COS7fibroblasts elevates uptake of long-chain fatty acids (Hui, T. Y. et al.(1998) J. Biol. Chem. 273:27420-27429).

[0009] Mitochondrial carrier proteins are transmembrane-spanningproteins which transport ions and charged metabolites between thecytosol and the mitochondrial matrix. Examples include the ADP, ATPcarrier protein; the 2-oxoglutarate/malate carrier; the phosphatecarrier protein; the pyruvate carrier; the dicarboxylate carrier whichtransports malate, succinate, fumarate, and phosphate; thetricarboxylate carrier which transports citrate and malate; and theGrave's disease carrier protein, a protein recognized by IgG in patientswith active Grave's disease, an autoimmune disorder resulting inhyperthyroidism. Proteins in this family consist of three tandem repeatsof an approximately 100 amino acid domain, each of which contains twotransmembrane regions (Stryer, L. (1995) Biochemistry, W. H. Freeman andCompany, New York N.Y., p. 551; PROSITE PDOC00189 Mitochondrial energytransfer proteins signature; Online Mendelian Inheritance in Man (OMIM)*275000 Graves Disease).

[0010] This class of transporters also includes the mitochondrialuncoupling proteins, which create proton leaks across the innermitochondrial membrane, thus uncoupling oxidative phosphorylation fromATP synthesis. The result is energy dissipation in the form of heatMitochondrial uncoupling proteins have been implicated as modulators ofthermoregulation and metabolic rate, and have been proposed as potentialtargets for drugs against metabolic diseases such as obesity (Ricquier,D. et al. (1999) J. Int. Med. 245:637-642).

[0011] Ion Channels

[0012] The electrical potential of a cell is generated and maintained bycontrolling the movement of ions across the plasma membrane. Themovement of ions requires ion channels, which form ion-selective poreswithin the membrane. There are two basic types of ion channels, iontransporters and gated ion channels. Ion transporters utilize the energyobtained from ATP hydrolysis to actively transport an ion against theion's concentration gradient. Gated ion channels allow passive flow ofan ion down the ion's electrochemical gradient under restrictedconditions. Together, these types of ion channels generate, maintain,and utilize an electrochemical gradient that is used in 1) electricalimpulse conduction down the axon of a nerve cell, 2) transport ofmolecules into cells against concentration gradients, 3) initiation ofmuscle contraction, and 4) endocrine cell secretion.

[0013] Ion Transporters

[0014] Ion transporters generate and maintain the resting electricalpotential of a cell. Utilizing the energy derived from ATP hydrolysis,they transport ions against the ion's concentration gradient. Thesetransmembrane ATPases are divided into three families. Thephosphorylated (P) class ion transporters, including Na⁺—K⁺ ATPase,Ca²⁺-ATPase, and H⁺-ATPase, are activated by a phosphorylation event.P-class ion transporters are responsible for maintaining restingpotential distributions such that cytosolic concentrations of Na⁺ andCa²⁺ are low and cytosolic concentration of K⁺ is high. The vacuolar (V)class of ion transporters includes H⁺ pumps on intracellular organelles,such as lysosomes and Golgi. V-class ion transporters are responsiblefor generating the low pH within the lumen of these organelles that isrequired for function. The coupling factor (F) class consists of H⁺pumps in the mitochondria. F-class ion transporters utilize a protongradient to generate ATP from ADP and inorganic phosphate (P_(i)).

[0015] The P-ATPases are hexamers of a 100 kD subunit with tentransmembrane domains and several large cytoplasmic regions that mayplay a role in ion binding (Scarborough, G. A. (1999) Curr. Opin. CellBiol. 11:517-522). The V-ATPases are composed of two functional domains:the V₁ domain, a peripheral complex responsible for ATP hydrolysis; andthe V₀ domain, an integral complex responsible for proton translocationacross the membrane. The F-ATPases are structurally and evolutionarilyrelated to the V-ATPases. The F-ATPase F₀ domain contains 12 copies ofthe c subunit, a highly hydrophobic protein composed of twotransmembrane domains and containing a single buried carboxyl group inTM2 that is essential for proton transport. The V-ATPase V₀ domaincontains three types of homologous c subunits with four or fivetransmembrane domains and the essential carboxyl group in TM4 or TM3.Both types of complex also contain a single a subunit that may beinvolved in regulating the pH dependence of activity (Forgac, M. (1999)J. Biol. Chem. 274:12951-12954).

[0016] The resting potential of the cell is utilized in many processesinvolving carrier proteins and gated ion channels. Carrier proteinsutilize the resting potential to transport molecules into and out of thecell. Amino acid and glucose transport into many cells is linked tosodium ion co-transport (symport) so that the movement of Na⁺ down anelectrochemical gradient drives transport of the other molecule up aconcentration gradient. Similarly, cardiac muscle links transfer of Ca²⁺out of the cell with transport of Na⁺ into the cell (antiport).

[0017] Gated Ion Channels

[0018] Gated ion channels control ion flow by regulating the opening andclosing of pores. The ability to control ion flux through various gatingmechanisms allows ion channels to mediate such diverse signaling andhomeostatic functions as neuronal and endocrine signaling, musclecontraction, fertilization, and regulation of ion and pH balance. Gatedion channels are categorized according to the manner of regulating thegating function. Mechanically-gated channels open their pores inresponse to mechanical stress; voltage-gated channels (e.g., Na⁺, K⁺,Ca²⁺, and Cl⁻ channels) open their pores in response to changes inmembrane potential; and ligand-gated channels (e.g., acetylcholine-,serotonin-, and glutamate-gated cation channels, and GABA- andglycine-gated chloride channels) open their pores in the presence of aspecific ion, nucleotide, or neurotransmitter. The gating properties ofa particular ion channel (i.e., its threshold for and duration ofopening and closing) are sometimes modulated by association withauxiliary channel proteins and/or post translational modifications, suchas phosphorylation.

[0019] Mechanically-gated or mechanosensitive ion channels act astransducers for the senses of touch, hearing, and balance, and also playimportant roles in cell volume regulation, smooth muscle contraction,and cardiac rhythm generation. A stretch-inactivated channel (SIC) wasrecently cloned from rat kidney. The SIC channel belongs to a group ofchannels which are activated by pressure or stress on the cell membraneand conduct both Ca²⁺ and Na⁺(Suzuki, M. et al. (1999) J. Biol. Chem.274:6330-6335).

[0020] The pore-forming subunits of the voltage-gated cation channelsform a superfamily of ion channel proteins. The characteristic domain ofthese channel proteins comprises six transmembrane domains (S1-S6), apore-forming region (P) located between S5 and S6, and intracellularamino and carboxy termini. In the Na⁺ and Ca²⁺ subfamilies, this domainis repeated four times, while in the K⁺ channel subfamily, each channelis formed from a tetramer of either identical or dissimilar subunits.The P region contains information specifying the ion selectivity for thechannel. In the case of K⁺ channels, a GYG tripeptide is involved inthis selectivity (Ishii, T. M. et al. (1997) Proc. Natl. Acad. Sci. USA94:11651-11656).

[0021] Voltage-gated Na⁺ and K⁺ channels are necessary for the functionof electrically excitable cells, such as nerve and muscle cells. Actionpotentials, which lead to neurotransmitter release and musclecontraction, arise from large, transient changes in the permeability ofthe membrane to Na⁺ and K⁺ ions. Depolarization of the membrane beyondthe threshold level opens voltage-gated Na⁺ channels. Sodium ions flowinto the cell, further depolarizing the membrane and opening morevoltage-gated Na⁺ channels, which propagates the depolarization down thelength of the cell. Depolarization also opens voltage-gated potassiumchannels. Consequently, potassium ions flow outward, which leads torepolarization of the membrane. Voltage-gated channels utilize chargedresidues in the fourth transmembrane segment (S4) to sense voltagechange. The open state lasts only about 1 millisecond, at which time thechannel spontaneously converts into an inactive state that cannot beopened irrespective of the membrane potential. Inactivation is mediatedby the channel's N-terminus, which acts as a plug that closes the pore.The transition from an inactive to a closed state requires a return toresting potential.

[0022] Voltage-gated Na⁺ channels are heterotrimeric complexes composedof a 260 kDa pore-forming α subunit that associates with two smallerauxiliary subunits, β1 and β2. The β2 subunit is a integral membraneglycoprotein that contains an extracellular Ig domain, and itsassociation with α and β1 subunits correlates with increased functionalexpression of the channel, a change in its gating properties, as well asan increase in whole cell capacitance due to an increase in membranesurface area (Isom, L. L. et al. (1995) Cell 83:433442).

[0023] Non voltage-gated Na⁺ channels include the members of theamiloride-sensitive Na⁺ channel/degenerin (NaC/DEG) family. Channelsubunits of this family are thought to consist of two transmembranedomains flanking a long extracellular loop, with the amino and carboxyltermini located within the cell. The NaC/DEG family includes theepithelial Na⁺ channel (ENAC) involved in Na⁺ reabsorption in epitheliaincluding the airway, distal colon, cortical collecting duct of thekidney, and exocrine duct glands. Mutations in ENaC result inpseudohypoaldosteronism type 1 and Liddle's syndrome(pseudohyperaldosteronism). The NaC/DEG family also includes therecently characterized H⁺-gated cation channels or acid-sensing ionchannels (ASIC). ASIC subunits are expressed in the brain and formheteromultimeric Na⁺-permeable channels. These channels require acid pHfluctuations for activation ASIC subunits show homology to thedegenerins, a family of mechanically-gated channels originally isolatedfrom C. elegans. Mutations in the degenerins cause neurodegeneration.ASIC subunits may also have a role in neuronal funtion, or in painperception, since tissue acidosis causes pain (Waldmann, R. and M.Lazdunski (1998) Curr. Opin. Neurobiol. 8:418424; Eglen, R. M. et al.(1999) Trends Pharmacol. Sci. 20:337-342).

[0024] K⁺ channels are located in all cell types, and may be regulatedby voltage, ATP concentration, or second messengers such as Ca²⁺ andcAMP. In non-excitable tissue, K⁺ channels are involved in proteinsynthesis, control of endocrine secretions, and the maintenance ofosmotic equilibrium across membranes. In neurons and other excitablecells, in addition to regulating action potentials and repolarizingmembranes, K⁺ channels are responsible for setting resting membranepotential. The cytosol contains non-diffusible anions and, to balancethis net negative charge, the cell contains a Na⁺—K⁺ pump and ionchannels that provide the redistribution of Na⁺, K⁺, and Cl⁻. The pumpactively transports Na⁺ out of the cell and K⁺ into the cell in a 3:2ratio. Ion channels in the plasma membrane allow K⁺ and Cl⁻ to flow bypassive diffusion. Because of the high negative charge within thecytosol, Cl⁻ flows out of the cell. The flow of K⁺ is balanced by anelectromotive force pulling K⁺ into the cell, and a K⁺ concentrationgradient pushing K⁺ out of the cell. Thus, the resting membranepotential is primarily regulated by K⁺ flow (Salkoff, L. and T. Jegla(1995) Neuron 15:489-492).

[0025] Potassium channel subunits of the Shaker-like superfamily allhave the characteristic six transmembrane/1 pore domain structure. Poursubunits combine as homo- or heterotetramers to form functional Kchannels. These pore-forming subunits also associate with variouscytoplasmic β subunits that alter channel inactivation kinetics. TheShaker-like channel family includes the voltage-gated K⁺ channels aswell as the delayed rectifier type channels such as the humanether-a-go-go related gene (HERG) associated with long QT, a cardiacdysrythmia syndrome (Curran, M. E. (1998) Curr. Opin. Biotechnol.9:565-572; Kaczarowski, G. J. and M. L. Garcia (1999) Curr. Opin. Chem.Biol. 3:448458).

[0026] A second superfamily of K⁺ channels is composed of the inwardrectifying channels (Kir). Kir channels have the property ofpreferentially conducting K⁺ currents in the inward direction. Theseproteins consist of a single potassium selective pore domain and twotransmembrane domains, which correspond to the fifth and sixthtransmembrane domains of voltage-gated K⁺ channels. Kir subunits alsoassociate as tetramers. The Kir family includes ROMK1, mutations inwhich lead to Bartter syndrome, a renal tubular disorder. Kir channelsare also involved in regulation of cardiac pacemaker activity, seizuresand epilepsy, and insulin regulation (Doupnik, C. A. et al. (1995) Curr.Opin. Neurobiol. 5:268-277; Curran, supra).

[0027] The recently recognized TWIKK⁺ channel family includes themammalian TWIK-1, TREK-1 and TASKproteins. Members of this familypossess an overall structure with four transmembrane domains and two Pdomains. These proteins are probably involved in controlling the restingpotential in a large set of cell types (Duprat, F. et al. (1997) EMBO J.16:5464-5471).

[0028] The voltage-gated Ca²⁺ channels have been classified into severalsubtypes based upon their electrophysiological and pharmacologicalcharacteristics. L-type Ca²⁺ channels are predominantly expressed inheart and skeletal muscle where they play an essential role inexcitation-contraction coupling. T-type channels are important forcardiac pacemaker activity, while N-type and P/Q-type channels areinvolved in the control of neurotransmitter release in the central andperipheral nervous system. The L-type and N-type voltage-gated Ca²⁺channels have been purified and, though their functions differdramatically, they have similar subunit compositions. The channels arecomposed of three subunits. The α₁ subunit forms the membrane pore andvoltage sensor, while the α₂δ and β subunits modulate thevoltage-dependence, gating properties, and the current amplitude of thechannel. These subunits are encoded by at least six α₁, one α₂δ, andfour β genes. A fourth subunit, γ, has been identified in skeletalmuscle (Walker, D. et al. (1998) J. Biol. Chem. 273:2361-2367;McCleskey, E. W. (1994) Curr. Opin. Neurobiol. 4:304-312).

[0029] Chloride channels are necessary in endocrine secretion and inregulation of cytosolic and organelle pH. In secretory epithelial cells,Cl⁻ enters the cell across a basolateral membrane through an Na⁺, K⁺/Cl⁻cotransporter, accumulating in the cell above its electrochemicalequilibrium concentration. Secretion of Cl⁻ from the apical surface, inresponse to hormonal stimulation, leads to flow of Na⁺ and water intothe secretory lumen. The cystic fibrosis transmembrane conductanceregulator (CFTR) is a chloride channel encoded by the gene for cysticfibrosis, a common fatal genetic disorder in humans. CFTR is a member ofthe ABC transporter family, and is composed of two domains eachconsisting of six transmembrane domains followed by a nucleotide-bindingsite. Loss of CFTR function decreases transepithelial water secretionand, as a result, the layers of mucus that coat the respiratory tree,pancreatic ducts, and intestine are dehydrated and difficult to clear.The resulting blockage of these sites leads to pancreatic insufficiency,“meconium ileus”, and devastating “chronic obstructive pulmonarydisease” (Al-Awqati, Q. et al. (1992) J. Exp. Biol. 172:245-266).

[0030] The voltage-gated chloride channels (CLC) are characterized by10-12 transmembrane domains, as well as two small globular domains knownas CBS domains. The CLC subunits probably function as homotetramers. CLCproteins are involved in regulation of cell volume, membrane potentialstabilization, signal transduction, and transepithelial transport.Mutations in CLC-1, expressed predominantly in skeletal muscle, areresponsible for autosomal recessive generalized myotonia and autosomaldominant myotonia congenita, while mutations in the kidney channel CLC-5lead to kidney stones (Jentsch, T. J. (1996) Curr. Opin. Neurobiol.6:303-310).

[0031] Ligand-gated channels open their pores when an extracellular orintracellular mediator binds to the channel. Neurotransmitter-gatedchannels are channels that open when a neurotransmitter binds to theirextracellular domain. These channels exist in the postsynaptic membraneof nerve or muscle cells. There are two types of neurotransmitter-gatedchannels. Sodium channels open in response to excitatoryneurotransmitters, such as acetylcholine, glutamate, and serotonin. Thisopening causes an influx of Na⁺ and produces the initial localizeddepolarization that activates the voltage-gated channels and starts theaction potential. Chloride channels open in response to inhibitoryneurotransmitters, such as γ-aminobutyric acid (GABA) and glycine,leading to hyperpolarization of the membrane and the subsequentgeneration of an action potential. Neurotransmitter-gated ion channelshave four transmembrane domains and probably function as pentamers(Jentsch, supra). Amino acids in the second transmembrane domain appearto be important in determining channel permeation and selectivity(Sather, W. A. et al. (1994) Curr. Opin. Neurobiol. 4:313-323).

[0032] Ligand-gated channels can be regulated by intracellular secondmessengers. For example, calcium-activated K⁺ channels are gated byinternal calcium ions. In nerve cells, an influx of calcium duringdepolarization opens K⁺ channels to modulate the magnitude of the actionpotential (Ishi et al., supra). The large conductance (BK) channel hasbeen purified from brain and its subunit composition determined. The αsubunit of the BK channel has seven rather than six transmembranedomains in contrast to voltage-gated K⁺ channels. The extratransmembrane domain is located at the subunit N-terminus. A28-amino-acid stretch in the C-terminal region of the subunit (the“calcium bowl” region) contains many negatively charged residues and isthought to be the region responsible for calcium binding. The β subunitconsists of two transmembrane domains connected by a glycosylatedextracellular loop, with intracellular N- and C-termini (Kaczorowski,supra; Vergara, C. et al. (1998) Curr. Opin. Neurobiol. 8:321-329).

[0033] Cyclic nucleotide-gated (CNG) channels are gated by cytosoliccyclic nucleotides. The best examples of these are the cAMP-gated Na⁺channels involved in olfaction and the cGMP-gated cation channelsinvolved in vision. Both systems involve ligand-mediated activation of aG-protein coupled receptor which then alters the level of cyclicnucleotide within the cell. CNG channels also represent a major pathwayfor Ca²⁺ entry into neurons, and play roles in neuronal development andplasticity. CNG channels are tetramers containing at least two types ofsubunits, an α subunit which can form functional homomeric channels, anda β subunit, which modulates the channel properties. All CNG subunitshave six transmembrane domains and a pore forming region between thefifth and sixth transmembrane domains, similar to voltage-gated K⁺channels. A large C-terminal domain contains a cyclic nucleotide bindingdomain, while the N-terminal domain confers variation among channelsubtypes (Zufall, F. et al. (1997) Curr. Opin. Neurobiol. 7.404412).

[0034] The activity of other types of ion channel proteins may also bemodulated by a variety of intracellular signalling proteins. Manychannels have sites for phosphorylation by one or more protein kinasesincluding protein kinase A, protein kinase C, tyrosine kinase, andcasein kinase II, all of which regulate ion channel activity in cells.Kir channels are activated by the binding of the Gβγ subunits ofheterotrimeric G-proteins (Reimann, F. and F. M. Ashcroft (1999) Curr.Opin. Cell. Biol. 11:503-508). Other proteins are involved in thelocalization of ion channels to specific sites in the cell membrane.Such proteins include the PDZ domain proteins known as MAGUKs(membrane-associated guanylate kinases) which regulate the clustering ofion channels at neuronal synapses (Craven, S. E. and D. S. Bredt (1998)Cell 93:495498).

[0035] Disease Correlation

[0036] The etiology of numerous human diseases and disorders can beattributed to defects in the transport of molecules across membranes.Defects in the trafficking of membrane-bound transporters and ionchannels are associated with several disorders, e.g., cystic fibrosis,glucose-galactose malabsorption syndrome, hypercholesterolemia, vonGierke disease, and certain forms of diabetes mellitus. Single-genedefect diseases resulting in an inability to transport small moleculesacross membranes include, e.g., cystinuria, iminoglycinuria, Hartupdisease, and Fanconi disease (van't Hoff, W. G. (1996) Exp. Nephrol.4:253-262; Talente, G. M. et al. (1994) Ann. Intern. Med. 120:218-226;and Chillon, M. et al. (1995) New Engl. J. Med. 332:1475-1480).

[0037] Human diseases caused by mutations in ion channel genes includedisorders of skeletal muscle, cardiac muscle, and the central nervoussystem. Mutations in the pore-forming subunits of sodium and chloridechannels cause myotonia, a muscle disorder in which relaxation aftervoluntary contraction is delayed. Sodium channel myotonias have beentreated with channel blockers. Mutations in muscle sodium and calciumchannels cause forms of periodic paralysis, while mutations in thesarcoplasmic calcium release channel, T-tubule calcium channel, andmuscle sodium channel cause malignant hyperthermia. Cardiac arrythmiadisorders such as the long QT syndromes and idiopathic ventricularfibrillation are caused by mutations in potassium and sodium channels(Cooper, E. C. and L. Y. Jan (1998) Proc. Natl. Acad. Sci. USA96:4759-4766). All four known human idiopathic epilepsy genes code forion channel proteins (Berkovic, S. F. and I. E. Scheffer (1999) Curr.Opin. Neurology 12:177-182). Other neurological disorders such asataxias, hemiplegic migraine and hereditary deafness can also resultfrom mutations in ion channel genes (Jen, J. (1999) Curr. Opin.Neurobiol. 9:274-280; Cooper, supra).

[0038] Ion channels have been the target for many drug therapies.Neurotransmitter-gated channels have been targeted in therapies fortreatment of insomnia, anxiety, depression, and schizophrenia.Voltage-gated channels have been targeted in therapies for arrhythmia,ischemic stroke, head trauma, and neurodegenerative disease (Taylor, C.P. and L. S. Narasimhan (1997) Adv. Pharmacol. 39:47-98). Variousclasses of ion channels also play an important role in the perception ofpain, and thus are potential targets for new analgesics. These includethe vanilloid-gated ion channels, which are activated by the vanilloidcapsaicin, as well as by noxious heat. Local anesthetics such aslidocaine and mexiletine which blockade voltage-gated Na⁺ channels havebeen useful in the treatment of neuropathic pain (Eglen, supra).

[0039] Ion channels in the immune system have recently been suggested astargets for immunomodulation. T-cell activation depends upon calciumsignaling, and a diverse set of T-cell specific ion channels has beencharacterized that affect this signaling process. Channel blockingagents can inhibit secretion of lymphokines, cell proliferation, andkilling of target cells. A peptide antagonist of the T-cell potassiumchannel Kv1.3 was found to suppress delayed-type hypersensitivity andallogenic responses in pigs, validating the idea of channel blockers assafe and efficacious immunosuppressants (Calahan, M. D. and K G. Chandy(1997) Curr. Opin. Biotechnol. 8:749-756).

[0040] The discovery of new transporters and ion channels and thepolynucleotides encoding them satisfies a need in the art by providingnew compositions which are useful in the diagnosis, prevention, andtreatment of transport, neurological, muscle, and immunologicaldisorders, and in the assessment of the effects of exogenous compoundson the expression of nucleic acid and amino acid sequences oftransporters and ion channels.

SUMMARY OF THE INVENTION

[0041] The invention features purified polypeptides, transporters andion channels, referred to collectively as “TRICH” and individually as“TRICH-1,” “TRICH-2,” “TRICH-3,” “TRICH-4,” “TRICH-5,” “TRICH-6,”“TRICH-7,” “TRICH-8,” “TRICH-9,” “TRICH-10,” “TRICH-11,” “TRICH-12,”“TRICH-13,” “TRICH-14,” “TRICH-15,” “TRICH-16,” “TRICH-17,” “TRICH-18,”“TRICH-19,” “TRICH-20,” “TRICH-21,” “TRICH-22,” “TRICH-23,” “TRICH-24,”“TRICH-25,” “TRICH-26,” and “TRICH-27.” In one aspect, the inventionprovides an isolated polypeptide comprising an amino acid sequenceselected from the group consisting of a) an amino acid sequence selectedfrom the group consisting of SEQ ID NO:1-27, b) a naturally occurringamino acid sequence having at least 90% sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:1-27, c) abiologically active fragment of an amino acid sequence selected from thegroup consisting of SEQ ID NO:1-27, and d) an immunogenic fragment of anamino acid sequence selected from the group consisting of SEQ ID NO:1-27. In one alternative, the invention provides an isolated polypeptidecomprising the amino acid sequence of SEQ ID NO:1-27.

[0042] The invention further provides an isolated polynucleotideencoding a polypeptide comprising an amino acid sequence selected fromthe group consisting of a) an amino acid sequence selected from thegroup consisting of SEQ ID NO:1-27, b) a naturally occurring amino acidsequence having at least 90% sequence identity to an amino acid sequenceselected from the group consisting of SEQ ID NO:1-27, c) a biologicallyactive fragment of an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27, and d) an immunogenic fragment of an aminoacid sequence selected from the group consisting of SEQ ID NO:1-27. Inone alternative, the polynucleotide encodes a polypeptide selected fromthe group consisting of SEQ ID NO:1-27. In another alternative, thepolynucleotide is selected from the group consisting of SEQ ID NO:28-54.

[0043] Additionally, the invention provides a recombinant polynucleotidecomprising a promoter sequence operably linked to a polynucleotideencoding a polypeptide comprising an amino acid sequence selected fromthe group consisting of a) an amino acid sequence selected from thegroup consisting of SEQ ID NO:1-27, b) a naturally occurring amino acidsequence having at least 90% sequence identity to an amino acid sequenceselected from the group consisting of SEQ ID NO:1-27, c) a biologicallyactive fragment of an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27, and d) an immunogenic fragment of an aminoacid sequence selected from the group consisting of SEQ ID NO:1-27. Inone alternative, the invention provides a cell transformed with therecombinant polynucleotide. In another alternative, the inventionprovides a transgenic organism comprising the recombinantpolynucleotide.

[0044] The invention also provides a method for producing a polypeptidecomprising an amino acid sequence selected from the group consisting ofa) an amino acid sequence selected from the group consisting of SEQ IDNO:1-27, b) a naturally occurring amino acid sequence having at least90% sequence identity to an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27, c) a biologically active fragment of anamino acid sequence selected from the group consisting of SEQ IDNO:1-27, and d) an immunogenic fragment of an amino acid sequenceselected from the group consisting of SEQ ID NO:1-27. The methodcomprises a) culturing a cell under conditions suitable for expressionof the polypeptide, wherein said cell is transformed with a recombinantpolynucleotide comprising a promoter sequence operably linked to apolynucleotide encoding the polypeptide, and b) recovering thepolypeptide so expressed.

[0045] Additionally, the invention provides an isolated antibody whichspecifically binds to a polypeptide comprising an amino acid sequenceselected from the group consisting of a) an amino acid sequence selectedfrom the group consisting of SEQ ID NO:1-27, b) a naturally occurringamino acid sequence having at least 90% sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:1-27, c) abiologically active fragment of an amino acid sequence selected from thegroup consisting of SEQ ID NO:1-27, and d) an immunogenic fragment of anamino acid sequence selected from the group consisting of SEQ IDNO:1-27.

[0046] The invention further provides an isolated polynucleotidecomprising a polynucleotide sequence selected from the group consistingof a) a polynucleotide sequence selected from the group consisting ofSEQ ID NO:28-54, b) a naturally occurring polynucleotide sequence havingat least 90% sequence identity to a polynucleotide sequence selectedfrom the group consisting of SEQ ID NO:28-54, c) a polynucleotidesequence complementary to a), d) a polynucleotide sequence complementaryto b), and e) an RNA equivalent of a)-d). In one alternative, thepolynucleotide comprises at least 60 contiguous nucleotides.

[0047] Additionally, the invention provides a method for detecting atarget polynucleotide in a sample, said target polynucleotide having asequence of a polynucleotide comprising a polynucleotide sequenceselected from the group consisting of a) a polynucleotide sequenceselected from the group consisting of SEQ ID NO:28-54, b) a naturallyoccurring polynucleotide sequence having at least 90% sequence identityto a polynucleotide sequence selected from the group consisting of SEQID NO:28-54, c) a polynucleotide sequence complementary to a), d) apolynucleotide sequence complementary to b), and e) an RNA equivalent ofa)-d). The method comprises a) hybridizing the sample with a probecomprising at least 20 contiguous nucleotides comprising a sequencecomplementary to said target polynucleotide in the sample, and whichprobe specifically hybridizes to said target polynucleotide, underconditions whereby a hybridization complex is formed between said probeand said target polynucleotide or fragments thereof, and b) detectingthe presence or absence of said hybridization complex, and optionally,if present, the amount thereof. In one alternative, the probe comprisesat least 60 contiguous nucleotides.

[0048] The invention further provides a method for detecting a targetpolynucleotide in a sample, said target polynucleotide having a sequenceof a polynucleotide comprising a polynucleotide sequence selected fromthe group consisting of a) a polynucleotide sequence selected from thegroup consisting of SEQ ID NO:28-54, b) a naturally occurringpolynucleotide sequence having at least 90% sequence identity to apolynucleotide sequence selected from the group consisting of SEQ IDNO:28-54, c) a polynucleotide sequence complementary to a), d) apolynucleotide sequence complementary to b), and e) an RNA equivalent ofa)-d). The method comprises a) amplifying said target polynucleotide orfragment thereof using polymerase chain reaction amplification, and b)detecting the presence or absence of said amplified targetpolynucleotide or fragment thereof, and, optionally, if present, theamount thereof. The invention further provides a composition comprisingan effective amount of a polypeptide comprising an amino acid sequenceselected from the group consisting of a) an amino acid sequence selectedfrom the group consisting of SEQ ID NO:1-27, b) a naturally occurringamino acid sequence having at least 90% sequence identity to an aminoacid sequence selected from the group consisting of SEQ ID NO:1-27, c) abiologically active fragment of an amino acid sequence selected from thegroup consisting of SEQ ID NO:1-27, and d) an immunogenic fragment of anamino acid sequence selected from the group consisting of SEQ IDNO:1-27, and a pharmaceutically acceptable excipient In one embodiment,the composition comprises an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27. The invention additionally provides amethod of treating a disease or condition associated with decreasedexpression of functional TRICH, comprising administering to a patient inneed of such treatment the composition.

[0049] The invention also provides a method for screening a compound foreffectiveness as an agonist of a polypeptide comprising an amino acidsequence selected from the group consisting of a) an amino acid sequenceselected from the group consisting of SEQ ID NO:1-27, b) a naturallyoccurring amino acid sequence having at least 90% sequence identity toan amino acid sequence selected from the group consisting of SEQ IDNO:1-27, c) a biologically active fragment of an amino acid sequenceselected from the group consisting of SEQ ID NO:1-27, and d) animmunogenic fragment of an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27. The method comprises a) exposing a samplecomprising the polypeptide to a compound, and b) detecting agonistactivity in the sample. In one alternative, the invention provides acomposition comprising an agonist compound identified by the method anda pharmaceutically acceptable excipient. In another alternative, theinvention provides a method of treating a disease or conditionassociated with decreased expression of functional TRICH, comprisingadministering to a patient in need of such treatment the composition.

[0050] Additionally, the invention provides a method for screening acompound for effectiveness as an antagonist of a polypeptide comprisingan amino acid sequence selected from the group consisting of a) an aminoacid sequence selected from the group consisting of SEQ ID NO:1-27, b) anaturally occurring amino acid sequence having at least 90% sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:1-27, c) a biologically active fragment of an amino acidsequence selected from the group consisting of SEQ ID NO:1-27, and d) animmunogenic fragment of an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27. The method comprises a) exposing a samplecomprising the polypeptide to a compound, and b) detecting antagonistactivity in the sample. In one alternative, the invention provides acomposition comprising an antagonist compound identified by the methodand a pharmaceutically acceptable excipient. In another alternative, theinvention provides a method of treating a disease or conditionassociated with overexpression of functional TRICH, comprisingadministering to a patient in need of such treatment the composition.

[0051] The invention further provides a method of screening for acompound that specifically binds to a polypeptide comprising an aminoacid sequence selected from the group consisting of a) an amino acidsequence selected from the group consisting of SEQ ID NO:1-27, b) anaturally occurring amino acid sequence having at least 90% sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:1-27, c) a biologically active fragment of an amino acidsequence selected from the group consisting of SEQ ID NO:1-27, and d) animmunogenic fragment of an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27. The method comprises a) combining thepolypeptide with at least one test compound under suitable conditions,and b) detecting binding of the polypeptide to the test compound,thereby identifying a compound that specifically binds to thepolypeptide.

[0052] The invention further provides a method of screening for acompound that modulates the activity of a polypeptide comprising anamino acid sequence selected from the group consisting of a) an aminoacid sequence selected from the group consisting of SEQ ID NO:1-27, b) anaturally occurring amino acid sequence having at least 90% sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO:1-27, c) a biologically active fragment of an amino acidsequence selected from the group consisting of SEQ ID NO:1-27, and d) animmunogenic fragment of an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27. The method comprises a) combining thepolypeptide with at least one test compound under conditions permissivefor the activity of the polypeptide, b) assessing the activity of thepolypeptide in the presence of the test compound, and c) comparing theactivity of the polypeptide in the presence of the test compound withthe activity of the polypeptide in the absence of the test compound,wherein a change in the activity of the polypeptide in the presence ofthe test compound is indicative of a compound that modulates theactivity of the polypeptide.

[0053] The invention further provides a method for screening a compoundfor effectiveness in altering expression of a target polynucleotide,wherein said target polynucleotide comprises a sequence selected fromthe group consisting of SEQ ID NO:28-54, the method comprising a)exposing a sample comprising the target polynucleotide to a compound,and b) detecting altered expression of the target polynucleotide.

[0054] The invention further provides a method for assessing toxicity ofa test compound, said method comprising a) treating a biological samplecontaining nucleic acids with the test compound; b) hybridizing thenucleic acids of the treated biological sample with a probe comprisingat least 20 contiguous nucleotides of a polynucleotide comprising apolynucleotide sequence selected from the group consisting of i) apolynucleotide sequence selected from the group consisting of SEQ IDNO:28-54, ii) a naturally occurring polynucleotide sequence having atleast 90% sequence identity to a polynucleotide sequence selected fromthe group consisting of SEQ ID NO:28-54, iii) a polynucleotide sequencecomplementary to i), iv) a polynucleotide sequence complementary to ii),and v) an RNA equivalent of i)-iv). Hybridization occurs underconditions whereby a specific hybridization complex is formed betweensaid probe and a target polynucleotide in the biological sample, saidtarget polynucleotide comprising a polynucleotide sequence selected fromthe group consisting of i) a polynucleotide sequence selected from thegroup consisting of SEQ ID NO:28-54, ii) a naturally occurringpolynucleotide sequence having at least 90% sequence identity to apolynucleotide sequence selected from the group consisting of SEQ IDNO:28-54, iii) a polynucleotide sequence complementary to i), iv) apolynucleotide sequence complementary to ii), and v) an RNA equivalentof i)-iv). Alternatively, the target polynucleotide comprises a fragmentof a polynucleotide sequence selected from the group consisting of i)-v)above; c) quantifying the amount of hybridization complex; and d)comparing the amount of hybridization complex in the treated biologicalsample with the amount of hybridization complex in an untreatedbiological sample, wherein a difference in the amount of hybridizationcomplex in the treated biological sample is indicative of toxicity ofthe test compound.

BRIEF DESCRIPTION OF THE TABLES

[0055] Table 1 summarizes the nomenclature for the full lengthpolynucleotide and polypeptide sequences of the present invention.

[0056] Table 2 shows the GenBank identification number and annotation ofthe nearest GenBank homolog for each polypeptide of the invention. Theprobability score for the match between each polypeptide and its GenBankhomolog is also shown.

[0057] Table 3 shows structural features of each polypeptide sequence,including predicted motifs and domains, along with the methods,algorithms, and searchable databases used for analysis of eachpolypeptide.

[0058] Table 4 lists the cDNA and genomic DNA fragments which were usedto assemble each polynucleotide sequence, along with selected fragmentsof the polynucleotide sequences.

[0059] Table 5 shows the representative cDNA library for eachpolynucleotide of the invention.

[0060] Table 6 provides an appendix which describes the tissues andvectors used for construction of the cDNA libraries shown in Table 5.

[0061] Table 7 shows the tools, programs, and algorithms used to analyzethe polynucleotides and polypeptides of the invention, along withapplicable descriptions, references, and threshold parameters.

DESCRIPTION OF THE INVENTION

[0062] Before the present proteins, nucleotide sequences, and methodsare described, it is understood that this invention is not limited tothe particular machines, materials and methods described, as these mayvary. It is also to be understood that the terminology used herein isfor the purpose of describing particular embodiments only, and is notintended to limit the scope of the present invention which will belimited only by the appended claims.

[0063] It must be noted that as used herein and in the appended claims,the singular forms “a,” “an,” and “the” include plural reference unlessthe context clearly dictates otherwise. Thus, for example, a referenceto “a host cell” includes a plurality of such host cells, and areference to “an antibody” is a reference to one or more antibodies andequivalents thereof known to those skilled in the art, and so forth.

[0064] Unless defined otherwise, all technical and scientific terms usedherein have the same meanings as commonly understood by one of ordinaryskill in the art to which this invention belongs. Although any machines,materials, and methods similar or equivalent to those described hereincan be used to practice or test the present invention, the preferredmachines, materials and methods are now described. All publicationsmentioned herein are cited for the purpose of describing and disclosingthe cell lines, protocols, reagents and vectors which are reported inthe publications and which might be used in connection with theinvention. Nothing herein is to be construed as an admission that theinvention is not entitled to antedate such disclosure by virtue of priorinvention.

[0065] Definitions

[0066] “TRICH” refers to the amino acid sequences of substantiallypurified TRICH obtained from any species, particularly a mammalianspecies, including bovine, ovine, porcine, murine, equine, and human,and from any source, whether natural, synthetic, semi-synthetic, orrecombinant.

[0067] The term “agonist” refers to a molecule which intensifies ormimics the biological activity of TRICH. Agonists may include proteins,nucleic acids, carbohydrates, small molecules, or any other compound orcomposition which modulates the activity of TRICH either by directlyinteracting with TRICH or by acting on components of the biologicalpathway in which TRICH participates.

[0068] An “allelic variant” is an alternative form of the gene encodingTRICH. Allelic variants may result from at least one mutation in thenucleic acid sequence and may result in altered mRNAs or in polypeptideswhose structure or function may or may not be altered. A gene may havenone, one, or many allelic variants of its naturally occurring form.Common mutational changes which give rise to allelic variants aregenerally ascribed to natural deletions, additions, or substitutions ofnucleotides. Each of these types of changes may occur alone, or incombination with the others, one or more times in a given sequence.

[0069] “Altered” nucleic acid sequences encoding TRICH include thosesequences with deletions, insertions, or substitutions of differentnucleotides, resulting in a polypeptide the same as TRICH or apolypeptide with at least one functional characteristic of TRICH.Included within this definition are polymorphisms which may or may notbe readily detectable using a particular oligonucleotide probe of thepolynucleotide encoding TRICH, and improper or unexpected hybridizationto allelic variants, with a locus other than the normal chromosomallocus for the polynucleotide sequence encoding TRICH. The encodedprotein may also be “altered,” and may contain deletions, insertions, orsubstitutions of amino acid residues which produce a silent change andresult in a functionally equivalent TRICH. Deliberate amino acidsubstitutions may be made on the basis of similarity in polarity,charge, solubility, hydrophobicity, hydrophilicity, and/or theamphipathic nature of the residues, as long as the biological orimmunological activity of TRICH is retained. For example, negativelycharged amino acids may include aspartic acid and glutamic acid, andpositively charged amino acids may include lysine and arginine. Aminoacids with uncharged polar side chains having similar hydrophilicityvalues may include: asparagine and glutamine; and serine and threonine.Amino acids with uncharged side chains having similar hydrophilicityvalues may include: leucine, isoleucine, and valine; glycine andalanine; and phenylalanine and tyrosine.

[0070] The terms “amino acid” and “amino acid sequence” refer to anoligopeptide, peptide, polypeptide, or protein sequence, or a fragmentof any of these, and to naturally occurring or synthetic molecules.Where “amino acid sequence” is recited to refer to a sequence of anaturally occurring protein molecule, “amino acid sequence” and liketerms are not meant to limit the amino acid sequence to the completenative amino acid sequence associated with the recited protein molecule.

[0071] “Amplification” relates to the production of additional copies ofa nucleic acid sequence. Amplification is generally carried out usingpolymerase chain reaction (PCR) technologies well known in the art.

[0072] The term “antagonist” refers to a molecule which inhibits orattenuates the biological activity of TRICH. Antagonists may includeproteins such as antibodies, nucleic acids, carbohydrates, smallmolecules, or any other compound or composition which modulates theactivity of TRICH either by directly interacting with TRICH or by actingon components of the biological pathway in which TRICH participates.

[0073] The term “antibody” refers to intact immunoglobulin molecules aswell as to fragments thereof, such as Fab, F(ab′)₂, and Fv fragments,which are capable of binding an epitopic determinant. Antibodies thatbind TRICH polypeptides can be prepared using intact polypeptides orusing fragments containing small peptides of interest as the immunizingantigen. The polypeptide or oligopeptide used to immunize an animal(e.g., a mouse, a rat, or a rabbit) can be derived from the translationof RNA, or synthesized chemically, and can be conjugated to a carrierprotein if desired. Commonly used carriers that are chemically coupledto peptides include bovine serum albumin, thyroglobulin, and keyholelimpet hemocyanin (KLH). The coupled peptide is then used to immunizethe animal.

[0074] The term “antigenic determinant” refers to that region of amolecule (i.e., an epitope) that makes contact with a particularantibody. When a protein or a fragment of a protein is used to immunizea host animal, numerous regions of the protein may induce the productionof antibodies which bind specifically to antigenic determinants(particular regions or three-dimensional structures on the protein). Anantigenic determinant may compete with the intact antigen (i.e., theimmunogen used to elicit the immune response) for binding to anantibody.

[0075] The term “antisense” refers to any composition capable ofbase-pairing with the “sense” (coding) strand of a specific nucleic acidsequence. Antisense compositions may include DNA; RNA; peptide nucleicacid (PNA); oligonucleotides having modified backbone linkages such asphosphorothioates, methylphosphonates, or benzylphosphonates;oligonucleotides having modified sugar groups such as 2′-methoxyethylsugars or 2′-methoxyethoxy sugars; or oligonucleotides having modifiedbases such as 5-methyl cytosine, 2′-deoxyuracil, or7-deaza-2′-deoxyguanosine. Antisense molecules may be produced by anymethod including chemical synthesis or transcription. Once introducedinto a cell, the complementary antisense molecule base-pairs with anaturally occurring nucleic acid sequence produced by the cell to formduplexes which block either transcription or translation. Thedesignation “negative” or “minus” can refer to the antisense strand, andthe designation “positive” or “plus” can refer to the sense strand of areference DNA molecule.

[0076] The term “biologically active” refers to a protein havingstructural, regulatory, or biochemical functions of a naturallyoccurring molecule. Likewise, “immunologically active” or “immunogenic”refers to the capability of the natural, recombinant, or syntheticTRICH, or of any oligopeptide thereof, to induce a specific immuneresponse in appropriate animals or cells and to bind with specificantibodies.

[0077] “Complementary” describes the relationship between twosingle-stranded nucleic acid sequences that anneal by base-pairing. Forexample, 5′-AGT-3′ pairs with its complement, 3′-TCA-5′.

[0078] A “composition comprising a given polynucleotide sequence” and a“composition comprising a given amino acid sequence” refer broadly toany composition containing the given polynucleotide or amino acidsequence. The composition may comprise a dry formulation or an aqueoussolution. Compositions comprising polynucleotide sequences encodingTRICH or fragments of TRICH may be employed as hybridization probes. Theprobes may be stored in freeze-dried form and may be associated with astabilizing agent such as a carbohydrate. In hybridizations, the probemay be deployed in an aqueous solution containing salts (e.g., NaCl),detergents (e.g., sodium dodecyl sulfate; SDS), and other components(e.g., Denhardt's solution, dry milk, salmon sperm DNA, etc.).

[0079] “Consensus sequence” refers to a nucleic acid sequence which hasbeen subjected to repeated DNA sequence analysis to resolve uncalledbases, extended using the XL-PCR kit (PE Biosystems, Foster City Calif.)in the 5′ and/or the 3′ direction, and resequenced, or which has beenassembled from one or more overlapping cDNA, EST, or genomic DNAfragments using a computer program for fragment assembly, such as theGEL VIEW fragment assembly system (GCG, Madison Wis.) or Phrap(University of Washington, Seattle Wash.). Some sequences have been bothextended and assembled to produce the consensus sequence.

[0080] “Conservative amino acid substitutions” are those substitutionsthat are predicted to least interfere with the properties of theoriginal protein, i.e., the structure and especially the function of theprotein is conserved and not significantly changed by suchsubstitutions. The table below shows amino acids which may besubstituted for an original amino acid in a protein and which areregarded as conservative amino acid substitutions. Original ResidueConservative Substitution Ala Gly, Ser Arg His, Lys Asn Asp, Gln, HisAsp Asn, Glu Cys Ala, Ser Gln Asn, Glu, His Glu Asp, Gln, His Gly AlaHis Asn, Arg, Gln, Glu Ile Leu, Val Leu Ile, Val Lys Arg, Gln, Glu MetLeu, Ile Phe His, Met, Leu, Trp, Tyr Ser Cys, Thr Thr Ser, Val Trp Phe,Tyr Tyr His, Phe, Trp Val Ile, Leu, Thr

[0081] Conservative amino acid substitutions generally maintain (a) thestructure of the polypeptide backbone in the area of the substitution,for example, as a beta sheet or alpha helical conformation, (b) thecharge or hydrophobicity of the molecule at the site of thesubstitution, and/or (c) the bulk of the side chain.

[0082] A “deletion” refers to a change in the amino acid or nucleotidesequence that results in the absence of one or more amino acid residuesor nucleotides.

[0083] The term “derivative” refers to a chemically modifiedpolynucleotide or polypeptide. Chemical modifications of apolynucleotide can include, for example, replacement of hydrogen by analkyl, acyl, hydroxyl, or amino group. A derivative polynucleotideencodes a polypeptide which retains at least one biological orimmunological function of the natural molecule. A derivative polypeptideis one modified by glycosylation, pegylation, or any similar processthat retains at least one biological or immunological function of thepolypeptide from which it was derived.

[0084] A “detectable label” refers to a reporter molecule or enzyme thatis capable of generating a measurable signal and is covalently ornoncovalently joined to a polynucleotide or polypeptide.

[0085] A “fragment” is a unique portion of TRICH or the polynucleotideencoding TRICH which is identical in sequence to but shorter in lengththan the parent sequence. A fragment may comprise up to the entirelength of the defined sequence, minus one nucleotide/amino acid residue.For example, a fragment may comprise from 5 to 1000 contiguousnucleotides or amino acid residues. A fragment used as a probe, primer,antigen, therapeutic molecule, or for other purposes, may be at least 5,10, 15, 16, 20, 25, 30, 40, 50, 60, 75, 100, 150, 250 or at least 500contiguous nucleotides or amino acid residues in length. Fragments maybe preferentially selected from certain regions of a molecule. Forexample, a polypeptide fragment may comprise a certain length ofcontiguous amino acids selected from the first 250 or 500 amino acids(or first 25% or 50%) of a polypeptide as shown in a certain definedsequence. Clearly these lengths are exemplary, and any length that issupported by the specification, including the Sequence Listing, tables,and figures, may be encompassed by the present embodiments.

[0086] A fragment of SEQ ID NO:28-54 comprises a region of uniquepolynucleotide sequence that specifically identifies SEQ ID NO:28-54,for example, as distinct from any other sequence in the genome fromwhich the fragment was obtained. A fragment of SEQ ID NO:28-54 isuseful, for example, in hybridization and amplification technologies andin analogous methods that distinguish SEQ ID NO:28-54 from relatedpolynucleotide sequences. The precise length of a fragment of SEQ IDNO:28-54 and the region of SEQ ID NO:28-54 to which the fragmentcorresponds are routinely determinable by one of ordinary skill in theart based on the intended purpose for the fragment.

[0087] A fragment of SEQ ID NO:1-27 is encoded by a fragment of SEQ IDNO:28-54. A fragment of SEQ ID NO: 1-27 comprises a region of uniqueamino acid sequence that specifically identifies SEQ ID NO:1-27. Forexample, a fragment of SEQ ID NO:1-27 is useful as an immunogenicpeptide for the development of antibodies that specifically recognizeSEQ ID NO:1-27. The precise length of a fragment of SEQ ID NO:1-27 andthe region of SEQ ID NO:1-27 to which the fragment corresponds areroutinely determinable by one of ordinary skill in the art based on theintended purpose for the fragment.

[0088] A “full length” polynucleotide sequence is one containing atleast a translation initiation codon (e.g., methionine) followed by anopen reading frame and a translation termination codon. A “full length”polynucleotide sequence encodes a “full length” polypeptide sequence.

[0089] “Homology” refers to sequence similarity or, interchangeably,sequence identity, between two or more polynucleotide sequences or twoor more polypeptide sequences.

[0090] The terms “percent identity” and “% identity,” as applied topolynucleotide sequences, refer to the percentage of residue matchesbetween at least two polynucleotide sequences aligned using astandardized algorithm. Such an algorithm may insert, in a standardizedand reproducible way, gaps in the sequences being compared in order tooptimize alignment between two sequences, and therefore achieve a moremeaningful comparison of the two sequences.

[0091] Percent identity between polynucleotide sequences may bedetermined using the default parameters of the CLUSTAL V algorithm asincorporated into the MEGALIGN version 3.12e sequence alignment program.This program is part of the LASERGENE software package, a suite ofmolecular biological analysis programs (DNASTAR, Madison Wis.). CLUSTALV is described in Higgins, D.G. and P. M. Sharp (1989) CABIOS 5:151-153and in Higgins, D. G. et al. (1992) CABIOS 8:189-191. For pairwisealignments of polynucleotide sequences, the default parameters are setas follows: Ktuple=2, gap penalty=5, window=4, and “diagonals saved”=4.The “weighted” residue weight table is selected as the default Percentidentity is reported by CLUSTAL V as the “percent similarity” betweenaligned polynucleotide sequences.

[0092] Alternatively, a suite of commonly used and freely availablesequence comparison algorithms is provided by the National Center forBiotechnology Information (NCBI) Basic Local Alignment Search Tool(BLAST) (Altschul, S. F. et al. (1990) J. Mol. Biol. 215:403410), whichis available from several sources, including the NCBI, Bethesda, Md.,and on the Internet at http://www.ncbi.nlmih.gov/BLAST/. The BLASTsoftware suite includes various sequence analysis programs including“blastn,” that is used to align a known polynucleotide sequence withother polynucleotide sequences from a variety of databases. Alsoavailable is a tool called “BLAST 2 Sequences” that is used for directpairwise comparison of two nucleotide sequences. “BLAST 2 Sequences” canbe accessed and used interactively athttp://www.ncbi.nlm.nihgov/gorf/bl2.html. The “BLAST 2 Sequences” toolcan be used for both blastn and blastp (discussed below). BLAST programsare commonly used with gap and other parameters set to default settings.For example, to compare two nucleotide sequences, one may use blastnwith the “BLAST 2 Sequences” tool Version 2.0.12 (April-21-2000) set atdefault parameters. Such default parameters may be, for example:

[0093] Matrix: BLOSUM62

[0094] Reward for match: 1

[0095] Penalty for mismatch: −2

[0096] Open Gap: 5 and Extension Gap: 2 penalties

[0097] Gap x drop-off: 50

[0098] Expect: 10

[0099] Word Size: 11

[0100] Filter: on

[0101] Percent identity may be measured over the length of an entiredefined sequence, for example, as defined by a particular SEQ ID number,or may be measured over a shorter length, for example, over the lengthof a fragment taken from a larger, defined sequence, for instance, afragment of at least 20, at least 30, at least 40, at least 50, at least70, at least 100, or at least 200 contiguous nucleotides. Such lengthsare exemplary only, and it is understood that any fragment lengthsupported by the sequences shown herein, in the tables, figures, orSequence Listing, may be used to describe a length over which percentageidentity may be measured.

[0102] Nucleic acid sequences that do not show a high degree of identitymay nevertheless encode similar amino acid sequences due to thedegeneracy of the genetic code. It is understood that changes in anucleic acid sequence can be made using this degeneracy to producemultiple nucleic acid sequences that all encode substantially the sameprotein.

[0103] The phrases “percent identity” and “% identity,” as applied topolypeptide sequences, refer to the percentage of residue matchesbetween at least two polypeptide sequences aligned using a standardizedalgorithm. Methods of polypeptide sequence alignment are well-known.Some alignment methods take into account conservative amino acidsubstitutions. Such conservative substitutions, explained in more detailabove, generally preserve the charge and hydrophobicity at the site ofsubstitution, thus preserving the structure (and therefore function) ofthe polypeptide.

[0104] Percent identity between polypeptide sequences may be determinedusing the default parameters of the CLUSTAL V algorithm as incorporatedinto the MEGALIGN version 3.12e sequence alignment program (describedand referenced above). For pairwise alignments of polypeptide sequencesusing CLUSTAL V, the default parameters are set as follows: Ktuple=1,gap penalty=3, window=5, and “diagonals saved”=5. The PAM250 matrix isselected as the default residue weight table. As with polynucleotidealignments, the percent identity is reported by CLUSTAL V as the“percent similarity” between aligned polypeptide sequence pairs.

[0105] Alternatively the NCBI BLAST software suite may be used. Forexample, for a pairwise comparison of two polypeptide sequences, one mayuse the “BLAST 2 Sequences” tool Version 2.0.12 (Apr.-21-2000) withblastp set at default parameters. Such default parameters may be, forexample:

[0106] Matrix: BLOSUM62

[0107] Open Gap: 11 and Extension Gap: 1 penalties

[0108] Gap x drop-off: 50

[0109] Expect: 10

[0110] Word Size: 3

[0111] Filter: on

[0112] Percent identity may be measured over the length of an entiredefined polypeptide sequence, for example, as defined by a particularSEQ ID number, or may be measured over a shorter length, for example,over the length of a fragment taken from a larger, defined polypeptidesequence, for instance, a fragment of at least 15, at least 20, at least30, at least 40, at least 50, at least 70 or at least 150 contiguousresidues. Such lengths are exemplary only, and it is understood that anyfragment length supported by the sequences shown herein, in the tables,figures or Sequence Listing, may be used to describe a length over whichpercentage identity may be measured.

[0113] “Human artificial chromosomes” (HACs) are linear microchromosomeswhich may contain DNA sequences of about 6 kb to 10 Mb in size and whichcontain all of the elements required for chromosome replication,segregation and maintenance.

[0114] The term “humanized antibody” refers to an antibody molecule inwhich the amino acid sequence in the non-antigen binding regions hasbeen altered so that the antibody more closely resembles a humanantibody, and still retains its original binding ability.

[0115] “Hybridization” refers to the process by which a polynucleotidestrand anneals with a complementary strand through base pairing underdefined hybridization conditions. Specific hybridization is anindication that two nucleic acid sequences share a high degree ofcomplementarity. Specific hybridization complexes form under permissiveannealing conditions and remain hybridized after the “washing” step(s).The washing step(s) is particularly important in determining thestringency of the hybridization process, with more stringent conditionsallowing less non-specific binding, i.e., binding between pairs ofnucleic acid strands that are not perfectly matched. Permissiveconditions for annealing of nucleic acid sequences are routinelydeterminable by one of ordinary skill in the art and may be consistentamong hybridization experiments, whereas wash conditions may be variedamong experiments to achieve the desired stringency, and thereforehybridization specificity. Permissive annealing conditions occur, forexample, at 68° C. in the presence of about 6×SSC, about 1% (w/v) SDS,and about 100 μg/ml sheared, denatured salmon sperm DNA.

[0116] Generally, stringency of hybridization is expressed, in part,with reference to the temperature under which the wash step is carriedout. Such wash temperatures are typically selected to be about 5° C. to20° C. lower than the thermal melting point (T_(m)) for the specificsequence at a defined ionic strength and pH. The T_(m) is thetemperature (under defined ionic strength and pH) at which 50% of thetarget sequence hybridizes to a perfectly matched probe. An equation forcalculating T_(m) and conditions for nucleic acid hybridization are wellknown and can be found in Sambrook, J. et al. (1989) Molecular Cloning:A Laboratory Manual, 2^(nd) ed., vol. 1-3, Cold Spring Harbor Press,Plainview N.Y.; specifically see volume 2, chapter 9.

[0117] High stringency conditions for hybridization betweenpolynucleotides of the present invention include wash conditions of 68°C. in the presence of about 0.2×SSC and about 0.1% SDS, for 1 hour.Alternatively, temperatures of about 65° C., 60° C., 55° C., or 42° C.may be used. SSC concentration may be varied from about 0.1 to 2×SSC,with SDS being present at about 0.1%. Typically, blocking reagents areused to block non-specific hybridization. Such blocking reagentsinclude, for instance, sheared and denatured salmon sperm DNA at about100-200 μg/ml. Organic solvent, such as formamide at a concentration ofabout 35-50% v/v, may also be used under particular circumstances, suchas for RNA:DNA hybridizations. Useful variations on these washconditions will be readily apparent to those of ordinary skill in theart. Hybridization, particularly under high stringency conditions, maybe suggestive of evolutionary similarity between the nucleotides. Suchsimilarity is strongly indicative of a similar role for the nucleotidesand their encoded polypeptides.

[0118] The term “hybridization complex” refers to a complex formedbetween two nucleic acid sequences by virtue of the formation ofhydrogen bonds between complementary bases. A hybridization complex maybe formed in solution (e.g., C₀t or R₀t analysis) or formed between onenucleic acid sequence present in solution and another nucleic acidsequence immobilized on a solid support (e.g., paper, membranes,filters, chips, pins or glass slides, or any other appropriate substrateto which cells or their nucleic acids have been fixed).

[0119] The words “insertion” and “addition” refer to changes in an aminoacid or nucleotide sequence resulting in the addition of one or moreamino acid residues or nucleotides, respectively.

[0120] “Immune response” can refer to conditions associated withinflammation, trauma, immune disorders, or infectious or geneticdisease, etc. These conditions can be characterized by expression ofvarious factors, e.g., cytokines, chemokines, and other signalingmolecules, which may affect cellular and systemic defense systems.

[0121] An “immunogenic fragment” is a polypeptide or oligopeptidefragment of TRICH which is capable of eliciting an immune response whenintroduced into a living organism, for example, a mammal. The term“immunogenic fragment” also includes any polypeptide or oligopeptidefragment of TRICH which is useful in any of the antibody productionmethods disclosed herein or known in the art.

[0122] The term “microarray” refers to an arrangement of a plurality ofpolynucleotides, polypeptides, or other chemical compounds on asubstrate.

[0123] The terms “element” and “array element” refer to apolynucleotide, polypeptide, or other chemical compound having a uniqueand defined position on a microarray.

[0124] The term “modulate” refers to a change in the activity of TRICH.For example, modulation may cause an increase or a decrease in proteinactivity, binding characteristics, or any other biological, functional,or immunological properties of TRICH.

[0125] The phrases “nucleic acid” and “nucleic acid sequence” refer to anucleotide, oligonucleotide, polynucleotide, or any fragment thereof.These phrases also refer to DNA or RNA of genomic or synthetic originwhich may be single-stranded or double-stranded and may represent thesense or the antisense strand, to peptide nucleic acid (PNA), or to anyDNA-like or RNA-like material.

[0126] “Operably linked” refers to the situation in which a firstnucleic acid sequence is placed in a functional relationship with asecond nucleic acid sequence. For instance, a promoter is operablylinked to a coding sequence if the promoter affects the transcription orexpression of the coding sequence. Operably linked DNA sequences may bein close proximity or contiguous and, where necessary to join twoprotein coding regions, in the same reading frame.

[0127] “Peptide nucleic acid” (PNA) refers to an antisense molecule oranti-gene agent which comprises an oligonucleotide of at least about 5nucleotides in length linked to a peptide backbone of amino acidresidues ending in lysine. The terminal lysine confers solubility to thecomposition. PNAs preferentially bind complementary single stranded DNAor RNA and stop transcript elongation, and may be pegylated to extendtheir lifespan in the cell.

[0128] “Post-translational modification” of an TRICH may involvelipidation, glycosylation, phosphorylation, acetylation, racemization,proteolytic cleavage, and other modifications known in the art. Theseprocesses may occur synthetically or biochemically. Biochemicalmodifications will vary by cell type depending on the enzymatic milieuof TRICH.

[0129] “Probe” refers to nucleic acid sequences encoding TRICH, theircomplements, or fragments thereof, which are used to detect identical,allelic or related nucleic acid sequences. Probes are isolatedoligonucleotides or polynucleotides attached to a detectable label orreporter molecule. Typical labels include radioactive isotopes, ligands,chemiluminescent agents, and enzymes. “Primers” are short nucleic acids,usually DNA oligonucleotides, which may be annealed to a targetpolynucleotide by complementary base-pairing. The primer may then beextended along the target DNA strand by a DNA polymerase enzyme. Primerpairs can be used for amplification (and identification) of a nucleicacid sequence, e.g., by the polymerase chain reaction (PCR).

[0130] Probes and primers as used in the present invention typicallycomprise at least 15 contiguous nucleotides of a known sequence. Inorder to enhance specificity, longer probes and primers may also beemployed, such as probes and primers that comprise at least 20, 25, 30,40, 50, 60, 70, 80, 90, 100, or at least 150 consecutive nucleotides ofthe disclosed nucleic acid sequences. Probes and primers may beconsiderably longer than these examples, and it is understood that anylength supported by the specification, including the tables, figures,and Sequence Listing, may be used.

[0131] Methods for preparing and using probes and primers are describedin the references, for example Sambrook, J. et al. (1989) MolecularCloning: A Laboratory Manual, 2^(nd) ed., vol. 1-3, Cold Spring HarborPress, Plainview N.Y.; Ausubel, F. M. et al. (1987) Current Protocols inMolecular Biology, Greene Publ. Assoc. & Wiley-Intersciences, New YorkN.Y.; Innis, M. et al. (1990) PCR Protocols A Guide to Methods andApplications, Academic Press, San Diego Calif. PCR primer pairs can bederived from a known sequence, for example, by using computer programsintended for that purpose such as Primer (Version 0.5, 1991, WhiteheadInstitute for Biomedical Research, Cambridge Mass.).

[0132] Oligonucleotides for use as primers are selected using softwareknown in the art for such purpose. For example, OLIGO 4.06 software isuseful for the selection of PCR primer pairs of up to 100 nucleotideseach, and for the analysis of oligonucleotides and largerpolynucleotides of up to 5,000 nucleotides from an input polynucleotidesequence of up to 32 kilobases. Similar primer selection programs haveincorporated additional features for expanded capabilities. For example,the PrimOU primer selection program (available to the public from theGenome Center at University of Texas South West Medical Center, DallasTex.) is capable of choosing specific primers from megabase sequencesand is thus useful for designing primers on a genome-wide scope. ThePrimer3 primer selection program (available to the public from theWhitehead Institute/MIT Center for Genome Research, Cambridge Mass.)allows the user to input a “mispriming library,” in which sequences toavoid as primer binding sites are user-specified Primer3 is useful, inparticular, for the selection of oligonucleotides for microarrays. (Thesource code for the latter two primer selection programs may also beobtained from their respective sources and modified to meet the user'sspecific needs.) The PrimeGen program (available to the public from theUK Human Genome Mapping Project Resource Centre, Cambridge UK) designsprimers based on multiple sequence alignments, thereby allowingselection of primers that hybridize to either the most conserved orleast conserved regions of aligned nucleic acid sequences. Hence, thisprogram is useful for identification of both unique and conservedoligonucleotides and polynucleotide fragments. The oligonucleotides andpolynucleotide fragments identified by any of the above selectionmethods are useful in hybridization technologies, for example, as PCR orsequencing primers, microarray elements, or specific probes to identifyfully or partially complementary polynucleotides in a sample of nucleicacids. Methods of oligonucleotide selection are not limited to thosedescribed above.

[0133] A “recombinant nucleic acid” is a sequence that is not naturallyoccurring or has a sequence that is made by an artificial combination oftwo or more otherwise separated segments of sequence. This artificialcombination is often accomplished by chemical synthesis or, morecommonly, by the artificial manipulation of isolated segments of nucleicacids, e.g., by genetic engineering techniques such as those describedin Sambrook, supra. The term recombinant includes nucleic acids thathave been altered solely by addition, substitution, or deletion of aportion of the nucleic acid. Frequently, a recombinant nucleic acid mayinclude a nucleic acid sequence operably linked to a promoter sequence.Such a recombinant nucleic acid may be part of a vector that is used,for example, to transform a cell.

[0134] Alternatively, such recombinant nucleic acids may be part of aviral vector, e.g., based on a vaccinia virus, that could be use tovaccinate a mammal wherein the recombinant nucleic acid is expressed,inducing a protective immunological response in the mammal.

[0135] A “regulatory element” refers to a nucleic acid sequence usuallyderived from untranslated regions of a gene and includes enhancers,promoters, introns, and 5′ and 3′ untranslated regions (UTRs).Regulatory elements interact with host or viral proteins which controltranscription, translation, or RNA stability.

[0136] “Reporter molecules” are chemical or biochemical moieties usedfor labeling a nucleic acid, amino acid, or antibody. Reporter moleculesinclude radionuclides; enzymes; fluorescent, chemiluminescent, orchromogenic agents; substrates; cofactors; inhibitors; magneticparticles; and other moieties known in the art.

[0137] An “RNA equivalent,” in reference to a DNA sequence, is composedof the same linear sequence of nucleotides as the reference DNA sequencewith the exception that all occurrences of the nitrogenous base thymineare replaced with uracil, and the sugar backbone is composed of riboseinstead of deoxyribose.

[0138] The term “sample” is used in its broadest sense. A samplesuspected of containing TRICH, nucleic acids encoding TRICH, orfragments thereof may comprise a bodily fluid; an extract from a cell,chromosome, organelle, or membrane isolated from a cell; a cell; genomicDNA, RNA, or cDNA, in solution or bound to a substrate; a tissue; atissue print; etc.

[0139] The terms “specific binding” and “specifically binding” refer tothat interaction between a protein or peptide and an agonist, anantibody, an antagonist, a small molecule, or any natural or syntheticbinding composition. The interaction is dependent upon the presence of aparticular structure of the protein, e.g., the antigenic determinant orepitope, recognized by the binding molecule. For example, if an antibodyis specific for epitope “A,” the presence of a polypeptide comprisingthe epitope A, or the presence of free unlabeled A, in a reactioncontaining free labeled A and the antibody will reduce the amount oflabeled A that binds to the antibody.

[0140] The term “substantially purified” refers to nucleic acid or aminoacid sequences that are removed from their natural environment and areisolated or separated, and are at least 60% free, preferably at least75% free, and most preferably at least 90% free from other componentswith which they are naturally associated.

[0141] A “substitution” refers to the replacement of one or more aminoacid residues or nucleotides by different amino acid residues ornucleotides, respectively.

[0142] “Substrate” refers to any suitable rigid or semi-rigid supportincluding membranes, filters, chips, slides, wafers, fibers, magnetic ornonmagnetic beads, gels, tubing, plates, polymers, microparticles andcapillaries. The substrate can have a variety of surface forms, such aswells, trenches, pins, channels and pores, to which polynucleotides orpolypeptides are bound.

[0143] A “transcript image” refers to the collective pattern of geneexpression by a particular cell type or tissue under given conditions ata given time.

[0144] “Transformation” describes a process by which exogenous DNA isintroduced into a recipient cell. Transformation may occur under naturalor artificial conditions according to various methods well known in theart, and may rely on any known method for the insertion of foreignnucleic acid sequences into a prokaryotic or eukaryotic host cell. Themethod for transformation is selected based on the type of host cellbeing transformed and may include, but is not limited to, bacteriophageor viral infection, electroporation, heat shock, lipofection, andparticle bombardment. The term “transformed cells” includes stablytransformed cells in which the inserted DNA is capable of replicationeither as an autonomously replicating plasmid or as part of the hostchromosome, as well as transiently transformed cells which express theinserted DNA or RNA for limited periods of time.

[0145] A “transgenic organism,” as used herein, is any organism,including but not limited to animals and plants, in which one or more ofthe cells of the organism contains heterologous nucleic acid introducedby way of human intervention, such as by transgenic techniques wellknown in the art. The nucleic acid is introduced into the cell, directlyor indirectly by introduction into a precursor of the cell, by way ofdeliberate genetic manipulation, such as by microinjection or byinfection with a recombinant virus. The term genetic manipulation doesnot include classical cross-breeding, or in vitro fertilization, butrather is directed to the introduction of a recombinant DNA molecule.The transgenic organisms contemplated in accordance with the presentinvention include bacteria, cyanobacteria, fungi, plants and animals.The isolated DNA of the present invention can be introduced into thehost by methods known in the art, for example infection, transfection,transformation or transconjugation. Techniques for transferring the DNAof the present invention into such organisms are widely known andprovided in references such as Sambrook et al. (1989), supra.

[0146] A “variant” of a particular nucleic acid sequence is defined as anucleic acid sequence having at least 40% sequence identity to theparticular nucleic acid sequence over a certain length of one of thenucleic acid sequences using blastn with the “BLAST 2 Sequences” toolVersion 2.0.9 (May-07-1999) set at default parameters. Such a pair ofnucleic acids may show, for example, at least 50%, at least 60%, atleast 70%, at least 80%, at least 85%, at least 90%, at least 95% or atleast 98% or greater sequence identity over a certain defined length. Avariant may be described as, for example, an “allelic” (as definedabove), “splice,” “species,” or “polymorphic” variant. A splice variantmay have significant identity to a reference molecule, but willgenerally have a greater or lesser number of polynucleotides due toalternative splicing of exons during mRNA processing. The correspondingpolypeptide may possess additional functional domains or lack domainsthat are present in the reference molecule. Species variants arepolynucleotide sequences that vary from one species to another. Theresulting polypeptides will generally have significant amino acididentity relative to each other. A polymorphic variant is a variation inthe polynucleotide sequence of a particular gene between individuals ofa given species. Polymorphic variants also may encompass “singlenucleotide polymorphisms” (SNPs) in which the polynucleotide sequencevaries by one nucleotide base. The presence of SNPs may be indicativeof, for example, a certain population, a disease state, or a propensityfor a disease state.

[0147] A “variant” of a particular polypeptide sequence is defined as apolypeptide sequence having at least 40% sequence identity to theparticular polypeptide sequence over a certain length of one of thepolypeptide sequences using blastp with the “BLAST 2 Sequences” toolVersion 2.0.9 (May-07-1999) set at default parameters. Such a pair ofpolypeptides may show, for example, at least 50%, at least 60%, at least70%, at least 80%, at least 90%, at least 95%, or at least 98% orgreater sequence identity over a certain defined length of one of thepolypeptides.

[0148] The Invention

[0149] The invention is based on the discovery of new human transportersand ion channels (TRICH), the polynucleotides encoding TRICH, and theuse of these compositions for the diagnosis, treatment, or prevention oftransport, neurological, muscle, and immunological disorders.

[0150] Table 1 summarizes the nomenclature for the full lengthpolynucleotide and polypeptide sequences of the invention. Eachpolynucleotide and its corresponding polypeptide are correlated to asingle Incyte project identification number (Incyte Project ID). Eachpolypeptide sequence is denoted by both a polypeptide sequenceidentification number (Polypeptide SEQ ID NO:) and an Incyte polypeptidesequence number (Incyte Polypeptide ID) as shown. Each polynucleotidesequence is denoted by both a polynucleotide sequence identificationnumber (Polynucleotide SEQ ID NO:) and an Incyte polynucleotideconsensus sequence number (Incyte Polynucleotide ID) as shown.

[0151] Table 2 shows sequences with homology to the polypeptides of theinvention as identified by BLAST analysis against the GenBank protein(genpept) database. Columns 1 and 2 show the polypeptide sequenceidentification number (Polypeptide SEQ ID NO:) and the correspondingIncyte polypeptide sequence number (Incyte Polypeptide ID) for eachpolypeptide of the invention. Column 3 shows the GenBank identificationnumber (Genbank ID NO:) of the nearest GenBank homolog. Column 4 showsthe probability score for the match between each polypeptide and itsGenBank homolog. Column 5 shows the annotation of the GenBank homologalong with relevant citations where applicable, all of which areexpressly incorporated by reference herein.

[0152] Table 3 shows various structural features of each of thepolypeptides of the invention Columns 1 and 2 show the polypeptidesequence identification number (SEQ ID NO:) and the corresponding Incytepolypeptide sequence number (Incyte Polypeptide ID) for each polypeptideof the invention. Column 3 shows the number of amino acid residues ineach polypeptide. Column 4 shows potential phosphorylation sites, andcolumn 5 shows potential glycosylation sites, as determined by theMOTIFS program of the GCG sequence analysis software package (GeneticsComputer Group, Madison Wis.). Column 6 shows amino acid residuescomprising signature sequences, domains, and motifs. Column 7 showsanalytical methods for protein structure/function analysis and in somecases, searchable databases to which the analytical methods wereapplied.

[0153] As shown in Table 4, the full length polynucleotide sequences ofthe present invention were assembled using cDNA sequences or coding(exon) sequences derived from genomic DNA, or any combination of thesetwo types of sequences. Columns 1 and 2 list the polynucleotide sequenceidentification number (Polynucleotide SEQ ID NO:) and the correspondingIncyte polynucleotide consensus sequence number (Incyte PolynucleotideID) for each polynucleotide of the invention. Column 3 shows the lengthof each polynucleotide sequence in basepairs. Column 4 lists fragmentsof the polynucleotide sequences which are useful, for example, inhybridization or amplification technologies that identify SEQ IDNO:28-54 or that distinguish between SEQ ID NO:28-54 and relatedpolynucleotide sequences. Column 5 shows identification numberscorresponding to cDNA sequences, coding sequences (exons) predicted fromgenomic DNA, and/or sequence assemblages comprised of both cDNA andgenomic DNA. These sequences were used to assemble the full lengthpolynucleotide sequences of the invention Columns 6 and 7 of Table 4show the nucleotide start (5′) and stop (3′) positions of the cDNA andgenomic sequences in column 5 relative to their respective full lengthsequences.

[0154] The identification numbers in Column 5 of Table 4 may referspecifically, for example, to Incyte cDNAs along with theircorresponding cDNA libraries. For example, 6813453H1 is theidentification number of an Incyte cDNA sequence, and ADRETUR01 is thecDNA library from which it is derived. Incyte cDNAs for which cDNAlibraries are not indicated were derived from pooled cDNA libraries(e.g., 70207988V1). Alternatively, the identification numbers in column5 may refer to GenBank cDNAs or ESTs (e.g., g1947104) which contributedto the assembly of the full length polynucleotide sequences.Alternatively, the identification numbers in column 5 may refer tocoding regions predicted by Genscan analysis of genomic DNA. Forexample, GNN.g6554406_(—)006 is the identification number of aGenscan-predicted coding sequence, with g6554406 being the GenBankidentification number of the sequence to which Genscan was applied. TheGenscan-predicted coding sequences may have been edited prior toassembly. (See Example IV.) Alternatively, the identification numbers incolumn 5 may refer to assemblages of both cDNA and Genscan-predictedexons brought together by an “exon stitching” algorithm (See Example V.)Alternatively, the identification numbers in column 5 may refer toassemblages of both cDNA and Genscan-predicted exons brought together byan “exon-stretching” algoritm (See Example V.) In some cases, IncytecDNA coverage redundant with the sequence coverage shown in column 5 wasobtained to confirm the final consensus polynucleotide sequence, but therelevant Incyte cDNA identification numbers are not shown.

[0155] Table 5 shows the representative cDNA libraries for those fulllength polynucleotide sequences which were assembled using Incyte cDNAsequences. The representative cDNA library is the Incyte cDNA librarywhich is most frequently represented by the Incyte cDNA sequences whichwere used to assemble and confirm the above polynucleotide sequences.The tissues and vectors which were used to construct the cDNA librariesshown in Table 5 are described in Table 6.

[0156] The invention also encompasses TRICH variants. A preferred TRICHvariant is one which has at least about 80%, or alternatively at leastabout 90%, or even at least about 95% amino acid sequence identity tothe TRICH amino acid sequence, and which contains at least onefunctional or structural characteristic of TRICH.

[0157] The invention also encompasses polynucleotides which encodeTRICH. In a particular embodiment, the invention encompasses apolynucleotide sequence comprising a sequence selected from the groupconsisting of SEQ ID NO:28-54, which encodes TRICH. The polynucleotidesequences of SEQ ID NO:28-54, as presented in the Sequence Listing,embrace the equivalent RNA sequences, wherein occurrences of thenitrogenous base thymine are replaced with uracil, and the sugarbackbone is composed of ribose instead of deoxyribose.

[0158] The invention also encompasses a variant of a polynucleotidesequence encoding TRICH. In particular, such a variant polynucleotidesequence will have at least about 70%, or alternatively at least about85%, or even at least about 95% polynucleotide sequence identity to thepolynucleotide sequence encoding TRICH. A particular aspect of theinvention encompasses a variant of a polynucleotide sequence comprisinga sequence selected from the group consisting of SEQ ID NO:28-54 whichhas at least about 70%, or alternatively at least about 85%, or even atleast about 95% polynucleotide sequence identity to a nucleic acidsequence selected from the group consisting of SEQ ID NO:28-54. Any oneof the polynucleotide variants described above can encode an amino acidsequence which contains at least one functional or structuralcharacteristic of TRICH.

[0159] It will be appreciated by those skilled in the art that as aresult of the degeneracy of the genetic code, a multitude ofpolynucleotide sequences encoding TRICH, some bearing minimal similarityto the polynucleotide sequences of any known and naturally occurringgene, may be produced. Thus, the invention contemplates each and everypossible variation of polynucleotide sequence that could be made byselecting combinations based on possible codon choices. Thesecombinations are made in accordance with the standard triplet geneticcode as applied to the polynucleotide sequence of naturally occurringTRICH, and all such variations are to be considered as beingspecifically disclosed.

[0160] Although nucleotide sequences which encode TRICH and its variantsare generally capable of hybridizing to the nucleotide sequence of thenaturally occurring TRICH under appropriately selected conditions ofstringency, it may be advantageous to produce nucleotide sequencesencoding TRICH or its derivatives possessing a substantially differentcodon usage, e.g., inclusion of non-naturally occurring codons. Codonsmay be selected to increase te rate at which expression of the peptideoccurs in a particular prokaryotic or eukaryotic host in accordance withthe frequency with which particular codons are utilized by the host.Other reasons for substantially altering the nucleotide sequenceencoding TRICH and its derivatives without altering the encoded aminoacid sequences include the production of RNA transcripts having moredesirable properties, such as a greater half-life, than transcriptsproduced from the naturally occurring sequence.

[0161] The invention also encompasses production of DNA sequences whichencode TRICH and TRICH derivatives, or fragments thereof, entirely bysynthetic chemistry. After production, the synthetic sequence may beinserted into any of the many available expression vectors and cellsystems using reagents well known in the art. Moreover, syntheticchemistry may be used to introduce mutations into a sequence encodingTRICH or any fragment thereof.

[0162] Also encompassed by the invention are polynucleotide sequencesthat are capable of hybridizing to the claimed polynucleotide sequences,and, in particular, to those shown in SEQ ID NO:28-54 and fragmentsthereof under various conditions of stringency. (See, e.g., Wahl, G. M.and S. L. Berger (1987) Methods Enzymol. 152:399-407; Kimmel, A. R.(1987) Methods Enzymol. 152:507-511.) Hybridization conditions,including annealing and wash conditions, are described in “Definitions.”

[0163] Methods for DNA sequencing are well known in the art and may beused to practice any of the embodiments of the invention. The methodsmay employ such enzymes as the Klenow fragment of DNA polymerase I,SEQUENASE (US Biochemical, Cleveland Ohio), Taq polymerase (PEBiosystems), thermostable T7 polymerase (Amersham Pharmacia Biotech,Piscataway N.J.), or combinations of polymerases and proofreadingexonucleases such as those found in the ELONGASE amplification system(Life Technologies, Gaithersburg Md.). Preferably, sequence preparationis automated with machines such as the MICROLAB 2200 liquid transfersystem (Hamilton, Reno Nev.), PTC200 thermal cycler (MJ Research,Watertown Mass.) and ABI CATALYST 800 thermal cycler (PE Biosystems).Sequencing is then carried out using either the ABI 373 or 377 DNAsequencing system (PE Biosystems), the MEGABACE 1000 DNA sequencingsystem (Molecular Dynamics, Sunnyvale Calif.), or other systems known inthe art. The resulting sequences are analyzed using a variety ofalgorithms which are well known in the art. (See, e.g., Ausubel, F. M.(1997) Short Protocols in Molecular Biology, John Wiley & Sons, New YorkN.Y., unit 7.7; Meyers, R. A. (1995) Molecular Biology andBiotechnology, Wiley VCH, New York N.Y., pp. 856-853.)

[0164] The nucleic acid sequences encoding TRICH may be extendedutilizing a partial nucleotide sequence and employing various PCR-basedmethods known in the art to detect upstream sequences, such as promotersand regulatory elements. For example, one method which may be employed,restriction-site PCR, uses universal and nested primers to amplifyunknown sequence from genomic DNA within a cloning vector. (See, e.g.,Sarkar, G. (1993) PCR Methods Applic. 2:318-322.) Another method,inverse PCR, uses primers that extend in divergent directions to amplifyunknown sequence from a circularized template. The template is derivedfrom restriction fragments comprising a known genomic locus andsurrounding sequences. (See, e.g., Triglia, T. et al. (1988) NucleicAcids Res. 16:8186.) A third method, capture PCR, involves PCRamplification of DNA fragments adjacent to known sequences in human andyeast artificial chromosome DNA. (See, e.g., Lagerstrom, M. et al.(1991) PCR Methods Applic. 1:111-119.) In this method, multiplerestriction enzyme digestions and ligations may be used to insert anengineered double-stranded sequence into a region of unknown sequencebefore performing PCR. Other methods which may be used to retrieveunknown sequences are known in the art (See, e.g., Parker, J. D. et al.(1991) Nucleic Acids Res. 19:3055-3060). Additionally, one may use PCR,nested primers, and PROMOTERFINDER libraries (Clontech, Palo AltoCalif.) to walk genomic DNA. This procedure avoids the need to screenlibraries and is useful in finding intron/exon junctions. For allPCR-based methods, primers may be designed using commercially availablesoftware, such as OLIGO 4.06 primer analysis software (NationalBiosciences, Plymouth Minn.) or another appropriate program, to be about22 to 30 nucleotides in length, to have a GC content of about 50% ormore, and to anneal to the template at temperatures of about 68° C. to72° C.

[0165] When screening for full length cDNAs, it is preferable to uselibraries that have been size-selected to include larger cDNAs. Inaddition, random-primed libraries, which often include sequencescontaining the 5′ regions of genes, are preferable for situations inwhich an oligo d(T) library does not yield a full-length cDNA. Genomiclibraries may be useful for extension of sequence into 5′non-transcribed regulatory regions.

[0166] Capillary electrophoresis systems which are commerciallyavailable may be used to analyze the size or confirm the nucleotidesequence of sequencing or PCR products. In particular, capillarysequencing may employ flowable polymers for electrophoretic separation,four different nucleotide-specific, laser-stimulated fluorescent dyes,and a charge coupled device camera for detection of the emittedwavelengths. Output/light intensity may be converted to electricalsignal using appropriate software (e.g., GENOTYPER and SEQUENCENAVIGATOR, PE Biosystems), and the entire process from loading ofsamples to computer analysis and electronic data display may be computercontrolled. Capillary electrophoresis is especially preferable forsequencing small DNA fragments which may be present in limited amountsin a particular sample.

[0167] In another embodiment of the invention, polynucleotide sequencesor fragments thereof which encode TRICH may be cloned in recombinant DNAmolecules that direct expression of TRICH, or fragments or functionalequivalents thereof, in appropriate host cells. Due to the inherentdegeneracy of the genetic code, other DNA sequences which encodesubstantially the same or a functionally equivalent amino acid sequencemay be produced and used to express TRICH.

[0168] The nucleotide sequences of the present invention can beengineered using methods generally known in the art in order to alterTRICH-encoding sequences for a variety of purposes including, but notlimited to, modification of the cloning, processing, and/or expressionof the gene product DNA shuffling by random fragmentation and PCRreassembly of gene fragments and synthetic oligonucleotides may be usedto engineer the nucleotide sequences. For example,oligonucleotide-mediated site-directed mutagenesis may be used tointroduce mutations that create new restriction sites, alterglycosylation patterns, change codon preference, produce splicevariants, and so forth.

[0169] The nucleotides of the present invention may be subjected to DNAshuffling techniques such as MOLECULARBREEDING (Maxygen Inc., SantaClara Calif.; described in U.S. Pat. No. 5,837,458; Chang, C. -C. et al.(1999) Nat. Biotechnol. 17:793-797; Christians, F. C. et al. (1999) Nat.Biotechnol. 17:259-264; and Crameri, A. et al. (1996) Nat Biotechnol.14:315-319) to alter or improve the biological properties of TRICH, suchas its biological or enzymatic activity or its ability to bind to othermolecules or compounds. DNA shuffling is a process by which a library ofgene variants is produced using PCR-mediated recombination of genefragments. The library is then subjected to selection or screeningprocedures that identify those gene variants with the desiredproperties. These preferred variants may then be pooled and furthersubjected to recursive rounds of DNA shuffling and selection/screening.Thus, genetic diversity is created through “artificial” breeding andrapid molecular evolution. For example, fragments of a single genecontaining random point mutations may be recombined, screened, and thenreshuffled until the desired properties are optimized. Alternatively,fragments of a given gene may be recombined with fragments of homologousgenes in the same gene family, either from the same or differentspecies, thereby maximizing the genetic diversity of multiple naturallyoccurring genes in a directed and controllable manner.

[0170] In another embodiment, sequences encoding TRICH may besynthesized, in whole or in part, using chemical methods well known inthe art (See, e.g., Caruthers, M. H. et al. (1980) Nucleic Acids Symp.Ser. 7:215-223; and Horn, T. et al. (1980) Nucleic Acids Symp. Ser.7:225-232.) Alternatively, TRICH itself or a fragment thereof may besynthesized using chemical methods. For example, peptide synthesis canbe performed using various solution-phase or solid-phase techniques.(See, e.g., Creighton, T. (1984) Proteins, Structures and MolecularProperties, W H Freeman, New York N.Y., pp. 55-60; and Roberge, J. Y. etal. (1995) Science 269:202-204.) Automated synthesis may be achievedusing the ABI 431A peptide synthesizer (PE Biosystems). Additionally,the amino acid sequence of TRICH, or any part thereof, may be alteredduring direct synthesis and/or combined with sequences from otherproteins, or any part thereof, to produce a variant polypeptide or apolypeptide having a sequence of a naturally occurring polypeptide.

[0171] The peptide may be substantially purified by preparative highperformance liquid chromatography. (See, e.g., Chiez, R. M. and F. Z.Regnier (1990) Methods Enzymol. 182:392-421.) The composition of thesynthetic peptides may be confirmed by amino acid analysis or bysequencing. (See, e.g., Creighton, supra, pp. 28-53.)

[0172] In order to express a biologically active TRICH, the nucleotidesequences encoding TRICH or derivatives thereof may be inserted into anappropriate expression vector, i.e., a vector which contains thenecessary elements for transcriptional and translational control of theinserted coding sequence in a suitable host. These elements includeregulatory sequences, such as enhancers, constitutive and induciblepromoters, and 5′ and 3′ untranslated regions in the vector and inpolynucleotide sequences encoding TRICH. Such elements may vary in theirstrength and specificity. Specific initiation signals may also be usedto achieve more efficient translation of sequences encoding TRICH. Suchsignals include the ATG initiation codon and adjacent sequences, e.g.the Kozak sequence. In cases where sequences encoding TRICH and itsinitiation codon and upstream regulatory sequences are inserted into theappropriate expression vector, no additional transcriptional ortranslational control signals may be needed. However, in cases whereonly coding sequence, or a fragment thereof, is inserted, exogenoustranslational control signals including an in-frame ATG initiation codonshould be provided by the vector. Exogenous translational elements andinitiation codons may be of various origins, both natural and synthetic.The efficiency of expression may be enhanced by the inclusion ofenhancers appropriate for the particular host cell system used (See,e.g., Scharf, D. et al. (1994) Results Probl. Cell Differ. 20:125-162.)

[0173] Methods-which are well known to those skilled in the art may beused to construct expression vectors containing sequences encoding TRICHand appropriate transcriptional and translational control elements.These methods include in vitro recombinant DNA techniques, synthetictechniques, and in vivo genetic recombination. (See, e.g., Sambrook, J.et al. (1989) Molecular Cloning, A Laboratory Manual, Cold Spring HarborPress, Plainview N.Y., ch. 4, 8, and 16-17; Ausubel, F. M. et al. (1995)Current Protocols in Molecular Biology, John Wiley & Sons, New YorkN.Y., ch. 9, 13, and 16.)

[0174] A variety of expression vector/host systems may be utilized tocontain and express sequences encoding TRICH. These include, but are notlimited to, microorganisms such as bacteria transformed with recombinantbacteriophage, plasmid, or cosmid DNA expression vectors; yeasttransformed with yeast expression vectors; insect cell systems infectedwith viral expression vectors (e.g., baculovirus); plant cell systemstransformed with viral expression vectors (e.g., cauliflower mosaicvirus, CaMV, or tobacco mosaic virus, TMV) or with bacterial expressionvectors (e.g., Ti or pBR322 plasmids); or animal cell systems. (See,e.g., Sambrook, supra; Ausubel, supra; Van Heeke, G. and S. M. Schuster(1989) J. Biol. Chem. 264:5503-5509; Engelhard, E. K. et al. (1994)Proc. Natl. Acad. Sci. USA 91:3224-3227; Sandig, V. et al. (1996) Hum.Gene Ther. 7:1937-1945; Takamatsu, N. (1987) EMBO J. 6:307-311; TheMcGraw Hill Yearbook of Science and Technology (1992) McGraw Hill, NewYork N.Y., pp. 191-196; Logan, J. and T. Shenk (1984) Proc. Natl. Acad.Sci. USA 81:3655-3659; and Harrington, J. J. et al. (1997) Nat. Genet.15:345-355.) Expression vectors derived from retroviruses, adenoviruses,or herpes or vaccinia viruses, or from various bacterial plasmids, maybe used for delivery of nucleotide sequences to the targeted organ,tissue, or cell population. (See, e.g., Di Nicola, M. et al. (1998)Cancer Gen. Ther. 5(6):350-356; Yu, M. et al. (1993) Proc. Natl. Acad.Sci. USA 90(13):6340-6344; Buller, R. M. et al. (1985) Nature317(6040):813-815; McGregor, D. P. et al. (1994) Mol. Immunol.31(3):219-226; and Verma, I. M. and N. Somia (1997) Nature 389:239-242.)The invention is not limited by the host cell employed.

[0175] In bacterial systems, a number of cloning and expression vectorsmay be selected depending upon the use intended for polynucleotidesequences encoding TRICH. For example, routine cloning, subcloning, andpropagation of polynucleotide sequences encoding TRICH can be achievedusing a multifunctional E. coli vector such as PBLUESCRIPT (Stratagene,La Jolla Calif.) or PSPORT1 plasmid (Life Technologies). Ligation ofsequences encoding TRICH into the vector's multiple cloning sitedisrupts the lacZ gene, allowing a calorimetric screening procedure foridentification of transformed bacteria containing recombinant molecules.In addition, these vectors may be useful for in vitro transcription,dideoxy sequencing, single strand rescue with helper phage, and creationof nested deletions in the cloned sequence. (See, e.g., Van Heeke, G.and S. M. Schuster (1989) J. Biol. Chem. 264:5503-5509.) When largequantities of TRICH are needed, e.g. for the production of antibodies,vectors which direct high level expression of TRICH may be used. Forexample, vectors containing the strong, inducible SP6 or T7bacteriophage promoter may be used.

[0176] Yeast expression systems may be used for production of TRICH. Anumber of vectors containing constitutive or inducible promoters, suchas alpha factor, alcohol oxidase, and PGH promoters, may be used in theyeast Saccharomyces cerevisiae or Pichia pastoris. In addition, suchvectors direct either the secretion or intracellular retention ofexpressed proteins and enable integration of foreign sequences into thehost genome for stable propagation. (See, e.g., Ausubel, 1995, supra;Bitter, G. A et al. (1987) Methods Enzymol. 153:516-544; and Scorer, C.A. et al. (1994) Bio/Technology 12:181-184.)

[0177] Plant systems may also be used for expression of TRICH.Transcription of sequences encoding TRICH may be driven by viralpromoters, e.g., the 35S and 19S promoters of CaMV used alone or incombination with the omega leader sequence from TMV (Takamatsu, N.(1987) EMBO J. 6:307-311). Alternatively, plant promoters such as thesmall subunit of RUBISCO or heat shock promoters may be used. (See,e.g., Coruzzi, G. et al. (1984) EMBO J. 3:1671-1680; Broglie, R. et al.(1984) Science 224:838-843; and Winter, J. et al. (1991) Results Probl.Cell Differ. 17:85-105.) These constructs can be introduced into plantcells by direct DNA transformation or pathogen-mediated transfection.(See, e.g., The McGraw Hill Yearbook of Science and Technology (1992)McGraw Hill, New York N.Y., pp. 191-196.)

[0178] In mammalian cells, a number of viral-based expression systemsmay be utilized. In cases where an adenovirus is used as an expressionvector, sequences encoding TRICH may be ligated into an adenovirustranscription/translation complex consisting of the late promoter andtripartite leader sequence. Insertion in a non-essential E1 or E3 regionof the viral genome may be used to obtain infective virus whichexpresses TRICH in host cells. (See, e.g., Logan, J. and T. Shenk (1984)Proc. Natl. Acad. Sci. USA 81:3655-3659.) In addition, transcriptionenhancers, such as the Rous sarcoma virus (RSV) enhancer, may be used toincrease expression in mammalian host cells. SV40 or EBV-based vectorsmay also be used for high-level protein expression.

[0179] Human artificial chromosomes (HACs) may also be employed todeliver larger fragments of DNA than can be contained in and expressedfrom a plasmid. HACs of about 6 kb to 10 Mb are constructed anddelivered via conventional delivery methods (liposomes, polycationicamino polymers, or vesicles) for therapeutic purposes. (See, e.g.,Harrington, J. J. et al. (1997) Nat. Genet. 15:345-355.)

[0180] For long term production of recombinant proteins in mammaliansystems, stable expression of TRICH in cell lines is preferred. Forexample, sequences encoding TRICH can be transformed into cell linesusing expression vectors which may contain viral origins of replicationand/or endogenous expression elements and a selectable marker gene onthe same or on a separate vector. Following the introduction of thevector, cells may be allowed to grow for about 1 to 2 days in enrichedmedia before being switched to selective media. The purpose of theselectable marker is to confer resistance to a selective agent, and itspresence allows growth and recovery of cells which successfully expressthe introduced sequences. Resistant clones of stably transformed cellsmay be propagated using tissue culture techniques appropriate to thecell type.

[0181] Any number of selection systems may be used to recovertransformed cell lines. These include, but are not limited to, theherpes simplex virus thymidine kinase and adeninephosphoribosyltransferase genes, for use in tk⁻ and apr⁻ cells,respectively. (See, e.g., Wigler, M. et al. (1977) Cell 11:223-232;Lowy, I. et al. (1980) Cell 22:817-823.) Also, antimetabolite,antibiotic, or herbicide resistance can be used as the basis forselection. For example, dhfr confers resistance to methotrexate; neoconfers resistance to the aminoglycosides neomycin and G418; and als andpat confer resistance to chlorsulfuron and phosphinotricinacetyltransferase, respectively. (See, e.g., Wigler, M. et al. (1980)Proc. Natl. Acad. Sci. USA 77:3567-3570; Colbere-Garapin, F. et al.(1981) J. Mol. Biol. 150:1-14.) Additional selectable genes have beendescribed, e.g., trpB and hisD, which alter cellular requirements formetabolites. (See, e.g., Hartman, S. C. and R. C. Mulligan (1988) Proc.Natl. Acad. Sci. USA 85:8047-8051.) Visible markers, e.g., anthocyanins,green fluorescent proteins (GFP; Clontech), β glucuronidase and itssubstrate β-glucuronide, or luciferase and its substrate luciferin maybe used. These markers can be used not only to identify transformants,but also to quantify the amount of transient or stable proteinexpression attributable to a specific vector system. (See, e.g., Rhodes,C. A. (1995) Methods Mol. Biol. 55:121-131.)

[0182] Although the presence/absence of marker gene expression suggeststhat the gene of interest is also present, the presence and expressionof the gene may need to be confirmed. For example, if the sequenceencoding TRICH is inserted within a marker gene sequence, transformedcells containing sequences encoding TRICH can be identified by theabsence of marker gene function. Alternatively, a marker gene can beplaced in tandem with a sequence encoding TRICH under the control of asingle promoter. Expression of the marker gene in response to inductionor selection usually indicates expression of the tandem gene as well.

[0183] In general, host cells that contain the nucleic acid sequenceencoding TRICH and that express TRICH may be identified by a variety ofprocedures known to those of skill in the art. These procedures include,but are not limited to, DNA-DNA or DNA-RNA hybridizations, PCRamplification, and protein bioassay or immunoassay techniques whichinclude membrane, solution, or chip based technologies for the detectionand/or quantification of nucleic acid or protein sequences.

[0184] Immunological methods for detecting and measuring the expressionof TRICH using either specific polyclonal or monoclonal antibodies areknown in the art. Examples of such techniques include enzyme-linkedimmunosorbent assays (ELISAs), radioimunoassays (RIAs), and fluorescenceactivated cell sorting (FACS). A two-site, monoclonal-based immunoassayutilizing monoclonal antibodies reactive to two non-interferng epitopeson TRICH is preferred, but a competitive binding assay may be employed.These and other assays are well known in the art. (See, e.g., Hampton,R. et al. (1990) Serological Methods, a Laboratory Manual, APS Press,St. Paul Minn., Sect. IV; Coigan, J. E. et al. (1997) Current Protocolsin Immunology, Greene Pub. Associates and Wiley-Interscience, New YorkN.Y.; and Pound, J. D. (1998) Immunochemical Protocols, Humana Press,Totowa N.J.)

[0185] A wide variety of labels and conjugation techniques are known bythose skilled in the art and may be used in various nucleic acid andamino acid assays. Means for producing labeled hybridization or PCRprobes for detecting sequences related to polynucleotides encoding TRICHinclude oligolabeling, nick translation, end-labeling, or PCRamplification using a labeled nucleotide. Alternatively, the sequencesencoding TRICH, or any fragments thereof, may be cloned into a vectorfor the production of an mRNA probe. Such vectors are known in the art,are commercially available, and may be used to synthesize RNA probes invitro by addition of an appropriate RNA polymerase such as T7, T3, orSP6 and labeled nucleotides. These procedures may be conducted using avariety of commercially available kits, such as those provided byAmersham Pharmacia Biotech, Promega (Madison Wis.), and US Biochemical.Suitable reporter molecules or labels which may be used for ease ofdetection include radionuclides, enzymes, fluorescent, chemiluminescent,or chromogenic agents, as well as substrates, cofactors, inhibitors,magnetic particles, and the like.

[0186] Host cells transformed with nucleotide sequences encoding TRICHmay be cultured under conditions suitable for the expression andrecovery of the protein from cell culture. The protein produced by atransformed cell may be secreted or retained intracellularly dependingon the sequence and/or the vector used. As will be understood by thoseof skill in the art, expression vectors containing polynucleotides whichencode TRICH may be designed to contain signal sequences which directsecretion of TRICH through a prokaryotic or eukaryotic cell membrane.

[0187] In addition, a host cell strain may be chosen for its ability tomodulate expression of the inserted sequences or to process theexpressed protein in the desired fashion. Such modifications of thepolypeptide include, but are not limited to, acetylation, carboxylation,glycosylation, phosphorylation, lipidation, and acylation.Post-translational processing which cleaves a “prepro” or “pro” form ofthe protein may also be used to specify protein targeting, folding,and/or activity. Different host cells which have specific cellularmachinery and charactristic mechanisms for post-translational activities(e.g., CHO, HeLa, MDCK, HEK293, and W138) are available from theAmerican Type Culture Collection (ATCC, Manassas Va.) and may be chosento ensure the correct modification and processing of the foreignprotein.

[0188] In another embodiment of the invention, natural, modified, orrecombinant nucleic acid sequences encoding TRICH may be ligated to aheterologous sequence resulting in translation of a fusion protein inany of the aforementioned host systems. For example, a chimeric TRICHprotein containing a heterologous moiety that can be recognized by acommercially available antibody may facilitate the screening of peptidelibraries for inhibitors of TRICH activity. Heterologous protein andpeptide moieties may also facilitate purification of fusion proteinsusing commercially available affinity matrices. Such moieties include,but are not limited to, glutathione S-transferase (GST), maltose bindingprotein (MBP), thioredoxin (Trx), calmodulin binding peptide (CBP),6-His, FLAG, c-myc, and hemagglutinin (HA). GST, MBP, Trx, CBP, and6-His enable purification of their cognate fusion proteins onimmobilized glutathione, maltose, phenylarsine oxide, calmodulin, andmetal-chelate resins, respectively. FLAG, c-myc, and hemagglutinin (HA)enable immunoaffinity purification of fusion proteins using commerciallyavailable monoclonal and polyclonal antibodies that specificallyrecognize these epitope tags. A fusion protein may also be engineered tocontain a proteolytic cleavage site located between the TRICH encodingsequence and the heterologous protein sequence, so that TRICH may becleaved away from the heterologous moiety following purification.Methods for fusion protein expression and purification are discussed inAusubel (1995, supra, ch. 10). A variety of commercially available kitsmay also be used to facilitate expression and purification of fusionproteins.

[0189] In a further embodiment of the invention, synthesis ofradiolabeled TRICH may be achieved in vitro using the TNT rabbitreticulocyte lysate or wheat germ extract system (Promega). Thesesystems couple transcription and translation of protein-coding sequencesoperably associated with the T7, T3, or SP6 promoters. Translation takesplace in the presence of a radiolabeled amino acid precursor, forexample, ³⁵S-methionine.

[0190] TRICH of the present invention or fragments thereof may be usedto screen for compounds that specifically bind to TRICH. At least oneand up to a plurality of test compounds may be screened for specificbinding to TRICH. Examples of test compounds include antibodies,oligonucleotides, proteins (e.g., receptors), or small molecules.

[0191] In one embodiment, the compound thus identified is closelyrelated to the natural ligand of TRICH, e.g., a ligand or fragmentthereof, a natural substrate, a structural or functional mimetic, or anatural binding partner. (See, e.g., Coligan, J. E. et al. (1991)Current Protocols in Immunology 1(2): Chapter 5.) Similarly, thecompound can be closely related to the natural receptor to which TRICHbinds, or to at least a fragment of the receptor, e.g., the ligandbinding site. In either case, the compound can be rationally designedusing known techniques. In one embodiment, screening for these compoundsinvolves producing appropriate cells which express TRICH, either as asecreted protein or on the cell membrane. Preferred cells include cellsfrom mammals, yeast, Drosophila, or E. coli. Cells expressing TRICH orcell membrane fractions which contain TRICH are then contacted with atest compound and binding, stimulation, or inhibition of activity ofeither TRICH or the compound is analyzed.

[0192] An assay may simply test binding of a test compound to thepolypeptide, wherein binding is detected by a fluorophore, radioisotope,enzyme conjugate, or other detectable label. For example, the assay maycomprise the steps of combining at least one test compound with TRICH,either in solution or affixed to a solid support, and detecting thebinding of TRICH to the compound. Alternatively, the assay may detect ormeasure binding of a test compound in the presence of a labeledcompetitor. Additionally, the assay may be carried out using cell-freepreparations, chemical libraries, or natural product mixtures, and thetest compound(s) may be free in solution or affixed to a solid support.

[0193] TRICH of the present invention or fragments thereof may be usedto screen for compounds that modulate the activity of TRICH. Suchcompounds may include agonists, antagonists, or partial or inverseagonists. In one embodiment, an assay is performed under conditionspermissive for TRICH activity, wherein TRICH is combined with at leastone test compound, and the activity of TRICH in the presence of a testcompound is compared with the activity of TRICH in the absence of thetest compound. A change in the activity of TRICH in the presence of thetest compound is indicative of a compound that modulates the activity ofTRICH. Alternatively, a test compound is combined with an in vitro orcell-free system comprising TRICH under conditions suitable for TRICHactivity, and the assay is performed. In either of these assays, a testcompound which modulates the activity of TRICH may do so indirectly andneed not come in direct contact with the test compound. At least one andup to a plurality of test compounds may be screened.

[0194] In another embodiment, polynucleotides encoding TRICH or theirmammalian homologs may be “knocked out” in an animal model system usinghomologous recombination in embryonic stem (ES) cells. Such techniquesare well known in the art and are useful for the generation of animalmodels of human disease. (See, e.g., U.S. Pat. No. 5,175,383 and U.S.Pat. No. 5,767,337.) For example, mouse ES cells, such as the mouse129/SvJ cell line, are derived from the early mouse embryo and grown inculture. The ES cells are transformed with a vector containing the geneof interest disrupted by a marker gene, e.g., the neomycinphosphotransferase gene (neo; Capecchi, M. R. (1989) Science244:1288-1292). The vector integrates into the corresponding region ofthe host genome by homologous recombination. Alternatively, homologousrecombination takes place using the Cre-loxP system to knockout a geneof interest in a tissue- or developmental stage-specific manner (Marth,J. D. (1996) Clin Invest. 97:1999-2002; Wagner, K. U. et al. (1997)Nucleic Acids Res. 25:4323-4330). Transformed ES cells are identifiedand microinjected into mouse cell blastocysts such as those from theC57BL/6 mouse strain. The blastocysts are surgically transferred topseudopregnant dams, and the resulting chimeric progeny are genotypedand bred to produce heterozygous or homozygous strains. Transgenicanimals thus generated may be tested with potential therapeutic or toxicagents.

[0195] Polynucleotides encoding TRICH may also be manipulated in vitroin ES cells derived from human blastocysts. Human ES cells have thepotential to differentiate into at least eight separate cell lineagesincluding endoderm, mesoderm, and ectodermal cell types. These celllineages differentiate into, for example, neural cells, hematopoieticlineages, and cardiomyocytes (Thomson, J. A. et al. (1998) Science282:1145-1147).

[0196] Polynucleotides encoding TRICH can also be used to create“knockin” humanized animals (pigs) or transgenic animals (mice or rats)to model human disease. With knockin technology, a region of apolynucleotide encoding TRICH is injected into animal ES cells, and theinjected sequence integrates into the animal cell genome. Transformedcells are injected into blastulae, and the blastulae are implanted asdescribed above. Transgenic progeny or inbred lines are studied andtreated with potential pharmaceutical agents to obtain information ontreatment of a human disease. Alternatively, a mammal inbred tooverexpress TRICH, e.g., by secreting TRICH in its milk, may also serveas a convenient source of that protein (Janne, J. et al. (1998)Biotechnol. Annu. Rev. 4:55-74).

[0197] Therapeutics

[0198] Chemical and structural similarity, e.g., in the context ofsequences and motifs, exists between regions of TRICH and transportersand ion channels. Therefore, TRICH appears to play a role in transport,neurological, muscle, and immunological disorders. In the treatment ofdisorders associated with increased TRICH expression or activity, it isdesirable to decrease the expression or activity of TRICH. In thetreatment of disorders associated with decreased TRICH expression oractivity, it is desirable to increase the expression or activity ofTRICH.

[0199] Therefore, in one embodiment, TRICH or a fragment or derivativethereof may be administered to a subject to treat or prevent a disorderassociated with decreased expression or activity of TRICH. Examples ofsuch disorders include, but are not limited to, a transport disordersuch as akinesia, amyotrophic lateral sclerosis, ataxia telangiectasia,cystic fibrosis, Becker's muscular dystrophy, Bell's palsy,Charcot-Marie Tooth disease, diabetes mellitus, diabetes insipidus,diabetic neuropathy, Duchenne muscular dystrophy, hyperkalemic periodicparalysis, normokalemic periodic paralysis, Parkinson's disease,malignant hyperthermia, multidrug resistance, myasthenia gravis,myotonic dystrophy, catatonia, tardive dyskinesia, dystonias, peripheralneuropathy, cerebral neoplasms, prostate cancer, cardiac disordersassociated with transport, e.g., angina, bradyarrythmia, tachyarrthmia,hypertension, Long QT syndrome, myocarditis, cardiomyopathy, nemalinemyopathy, centronuclear myopathy, lipid myopathy, mitochondrialmyopathy, thyrotoxic myopathy, ethanol myopathy, dermatomyositis,inclusion body myositis, infectious myositis, polymyositis, neurologicaldisorders associated with transport, e.g., Alzheimer's disease, amnesia,bipolar disorder, dementia, depression, epilepsy, Tourette's disorder,paranoid psychoses, and schizophrenia, and other disorders associatedwith transport, e.g., neurofibromatosis, postherpetic neuralgia,trigeminal neuropathy, sarcoidosis, sickle cell anemia, Wilson'sdisease, cataracts, infertility, pulmonary artery stenosis,sensorineural autosomal deafness, hyperglycemia, hypoglycemia, Grave'sdisease, goiter, Cushing's disease, Addison's disease, glucose-galactosemalabsorption syndrome, hypercholesterolemia, adrenoleukodystrophy,Zellweger syndrome, Menkes disease, occipital horn syndrome, von Gierkedisease, cystinuria, iminoglycinuria, Hartup disease, and Fanconidisease; a neurological disorder such as epilepsy, ischemiccerebrovascular disease, stroke, cerebral neoplasms, Alzheimer'sdisease, Pick's disease, Huntington's disease, dementia, Parkinson'sdisease and other extrapyramidal disorders, amyotrophic lateralsclerosis and other motor neuron disorders, progressive neural muscularatrophy, retinitis pigmentosa, hereditary ataxias, multiple sclerosisand other demyelinating diseases, bacterial and viral meningitis, brainabscess, subdural empyema, epidural abscess, suppurative intracranialthrombophlebitis, myelitis and radiculitis, viral central nervous systemdisease, prion diseases including kuru, Creutzfeldt-Jakob disease, andGerstmann-Straussler-Scheinker syndrome, fatal familial insomnia,nutritional and metabolic diseases of the nervous system,neurofibromatosis, tuberous sclerosis, cerebelloretinalhemangioblastomatosis, encephalotrigeminal syndrome, mental retardationand other developmental disorders of the central nervous systemincluding Down syndrome, cerebral palsy, neuroskeletal disorders,autonomic nervous system disorders, cranial nerve disorders, spinal corddiseases, muscular dystrophy and other neuromuscular disorders,peripheral nervous system disorders, dermatomyositis and polymyositis,inherited, metabolic, endocrine, and toxic myopathies, myastheniagravis, periodic paralysis, mental disorders including mood, anxiety,and schizophrenic disorders, seasonal affective disorder (SAD),akathesia, amnesia, catatonia, diabetic neuropathy, tardive dyskinesia,dystonias, paranoid psychoses, postherpetic neuralgia, Tourette'sdisorder, progressive supranuclear palsy, corticobasal degeneration, andfamilial frontotemporal dementia; a muscle disorder such ascardiomyopathy, myocarditis, Duchenne's muscular dystrophy, Becker'smuscular dystrophy, myotonic dystrophy, central core disease, nemalinemyopathy, centronuclear myopathy, lipid myopathy, mitochondrialmyopathy, infectious myositis, polymyositis, dermatomyositis, inclusionbody myositis, thyrotoxic myopathy, ethanol myopathy, angina,anaphylactic shock, arrhythmias, asthma, cardiovascular shock, Cushing'ssyndrome, hypertension, hypoglycemia, myocardial infarction, migraine,pheochromocytoma, and myopathies including encephalopathy, epilepsy,Kearns-Sayre syndrome, lactic acidosis, myoclonic disorder,ophthalmoplegia, and acid maltase deficiency (AMD, also known as Pompe'sdisease); and an immunological disorder such as acquiredimmunodeficiency syndrome (AIDS), Addison's disease, adult respiratorydistress syndrome, allergies, ankylosing spondylitis, amyloidosis,anemia, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmunethyroiditis, autoimmune polyendocrinopathy-candidiasis-ectodermaldystrophy (APECED), bronchitis, cholecystitis, contact dermatitis,Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus,emphysema, episodic lymphopenia with lymphocytotoxins, erythroblastosisfetalis, erythema nodosum, atrophic gastritis, glomerulonephritis,Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis,hypereosinophilia, irritable bowel syndrome, multiple sclerosis,myasthenia gravis, myocardial or pericardial inflammation,osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis,Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjögren'ssyndrome, systemic anaphylaxis, systemic lupus erythematosus, systemicsclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Wernersyndrome, complications of cancer, hemodialysis, and extracorporealcirculation, viral, bacterial, fungal, parasitic, protozoal, andhelminthic infections, and trauma.

[0200] In another embodiment, a vector capable of expressing TRICH or afragment or derivative thereof may be administered to a subject to treator prevent a disorder associated with decreased expression or activityof TRICH including, but not limited to, those described above.

[0201] In a further embodiment, a composition comprising a substantiallypurified TRICH in conjunction with a suitable pharmaceutical carrier maybe administered to a subject to treat or prevent a disorder associatedwith decreased expression or activity of TRICH including, but notlimited to, those provided above.

[0202] In still another embodiment, an agonist which modulates theactivity of TRICH may be administered to a subject to treat or prevent adisorder associated with decreased expression or activity of TRICHincluding, but not limited to, those listed above.

[0203] In a further embodiment, an antagonist of TRICH may beadministered to a subject to treat or prevent a disorder associated withincreased expression or activity of TRICH. Examples of such disordersinclude, but are not limited to, those transport, neurological, muscle,and immunological disorders described above. In one aspect, an antibodywhich specifically binds TRICH may be used directly as an antagonist orindirectly as a targeting or delivery mechanism for bringing apharmaceutical agent to cells or tissues which express TRICH.

[0204] In an additional embodiment, a vector expressing the complementof the polynucleotide encoding TRICH may be administered to a subject totreat or prevent a disorder associated with increased expression oractivity of TRICH including, but not limited to, those described above.

[0205] In other embodiments, any of the proteins, antagonists,antibodies, agonists, complementary sequences, or vectors of theinvention may be administered in combination with other appropriatetherapeutic agents. Selection of the appropriate agents for use incombination therapy may be made by one of ordinary skill in the art,according to conventional pharmaceutical principles. The combination oftherapeutic agents may act synergistically to effect the treatment orprevention of the various disorders described above. Using thisapproach, one may be able to achieve therapeutic efficacy with lowerdosages of each agent, thus reducing the potential for adverse sideeffects.

[0206] An antagonist of TRICH may be produced using methods which aregenerally known in the art. In particular, purified TRICH may be used toproduce antibodies or to screen libraries of pharmaceutical agents toidentify those which specifically bind TRICH. Antibodies to TRICH mayalso be generated using methods that are well known in the art. Suchantibodies may include, but are not limited to, polyclonal, monoclonal,chimeric, and single chain antibodies, Fab fragments, and fragmentsproduced by a Fab expression library. Neutralizing antibodies (i.e.,those which inhibit dimer formation) are generally preferred fortherapeutic use.

[0207] For the production of antibodies, various hosts including goats,rabbits, rats, mice, humans, and others may be immunized by injectionwith TRICH or with any fragment or oligopeptide thereof which hasimmunogenic properties. Depending on the host species, various adjuvantsmay be used to increase immunological response. Such adjuvants include,but are not limited to, Freund's, mineral gels such as aluminumhydroxide, and surface active substances such as lysolecithin, pluronicpolyols, polyanions, peptides, oil emulsions, KLH, and dinitrophenol.Among adjuvants used in humans, BCG (bacili Calmette-Guerin) andCorynebacterium parvum are especially preferable.

[0208] It is preferred that the oligopeptides, peptides, or fragmentsused to induce antibodies to TRICH have an amino acid sequenceconsisting of at least about 5 amino acids, and generally will consistof at least about 10 amino acids. It is also preferable that theseoligopeptides, peptides, or fragments are identical to a portion of theamino acid sequence of the natural protein. Short stretches of TRICHamino acids may be fused with those of another protein, such as KLH, andantibodies to the chimeric molecule may be produced.

[0209] Monoclonal antibodies to TRICH may be prepared using anytechnique which provides for the production of antibody molecules bycontinuous cell lines in culture. These include, but are not limited to,the hybridoma technique, the human B-cell hybridoma technique, and theEBV-hybridoma technique. (See, e.g., Kohler, G. et al. (1975) Nature256:495-497; Kozbor, D. et al. (1985) J. Immunol. Methods 81:31-42;Cote, R. J. et al. (1983) Proc. Natl. Acad. Sci. USA 80:2026-2030; andCole, S. P. et al. (1984) Mol. Cell Biol. 62:109-120.)

[0210] In addition, techniques developed for the production of “chimericantibodies,” such as the splicing of mouse antibody genes to humanantibody genes to obtain a molecule with appropriate antigen specificityand biological activity, can be used. (See, e.g., Morrison, S. L. et al.(1984) Proc. Natl. Acad. Sci. USA 81:6851-6855; Neuberger, M. S. et al.(1984) Nature 312:604-608; and Takeda, S. et al. (1985) Nature314:452454.) Alternatively, techniques described for the production ofsingle chain antibodies may be adapted, using methods known in the art,to produce TRICH-specific single chain antibodies. Antibodies withrelated specificity, but of distinct idiotypic composition, may begenerated by chain shuffling from random combinatorial immunoglobulinlibraries. (See, e.g., Burton, D. R. (1991) Proc. Natl. Acad. Sci. USA88:10134-10137.)

[0211] Antibodies may also be produced by inducing in vivo production inthe lymphocyte population or by screening immunoglobulin libraries orpanels of highly specific binding reagents as disclosed in theliterature. (See, e.g., Orlandi, R. et al. (1989) Proc. Natl. Acad. Sci.USA 86:3833-3837; Winter, G. et al. (1991) Nature 349:293-299.)

[0212] Antibody fragments which contain specific binding sites for TRICHmay also be generated. For example, such fragments include, but are notlimited to, F(ab′)₂ fragments produced by pepsin digestion of theantibody molecule and Fab fragments generated by reducing the disulfidebridges of the F(ab′)2 fragments. Alternatively, Fab expressionlibraries may be constructed to allow rapid and easy identification ofmonoclonal Fab fragments with the desired specificity. (See, e.g., Huse,W. D. et al. (1989) Science 246:1275-1281.)

[0213] Various immunoassays may be used for screening to identifyantibodies having the desired specificity. Numerous protocols forcompetitive binding or immunoradiometric assays using either polyclonalor monoclonal antibodies with established specificities are well knownin the art. Such immunoassays typically involve the measurement ofcomplex formation between TRICH and its specific antibody. A two-site,monoclonal-based immunoassay utilizing monoclonal antibodies reactive totwo non-interfering TRICH epitopes is generally used, but a competitivebinding assay may also be employed (Pound, supra).

[0214] Various methods such as Scatchard analysis in conjunction withradioimmunoassay techniques may be used to assess the affinity ofantibodies for TRICH. Affinity is expressed as an association constant,K_(a), which is defined as the molar concentration of TRICH-antibodycomplex divided by the molar concentrations of free antigen and freeantibody under equilibrium conditions. The K_(a) determined for apreparation of polyclonal antibodies, which are heterogeneous in theiraffinities for multiple TRICH epitopes, represents the average affinity,or avidity, of the antibodies for TRICH. The K_(a) determined for apreparation of monoclonal antibodies, which are monospecific for aparticular TRICH epitope, represents a true measure of affinity.High-affinity antibody preparations with K_(a) ranging from about 10⁹ to10¹² L/mole are preferred for use in immunoassays in which theTRICH-antibody complex must withstand rigorous manipulations.Low-affinity antibody preparations with K_(a) ranging from about 10⁶ to10⁷ L/mole are preferred for use in immunopurification and similarprocedures which ultimately require dissociation of TRICH, preferably inactive form, from the antibody (Catty, D. (1988) Antibodies, Volume I: APractical Approach, IRL Press, Washington D.C.; Liddell, J. E. and A.Cryer (1991) A Practical Guide to Monoclonal Antibodies, John Wiley &Sons, New York N.Y.).

[0215] The titer and avidity of polyclonal antibody preparations may befurther evaluated to determine the quality and suitability of suchpreparations for certain downstream applications. For example, apolyclonal antibody preparation containing at least 1-2 mg specificantibody/ml, preferably 5-10 mg specific antibody/ml, is generallyemployed in procedures respiring precipitation of TRICH-antibodycomplexes. Procedures for evaluating antibody specificity, titer, andavidity, and guidelines for antibody quality and usage in variousapplications, are generally available. (See, e.g., Catty, supra, andColigan et al. supra.)

[0216] In another embodiment of the invention, the polynucleotidesencoding TRICH, or any fragment or complement thereof, may be used fortherapeutic purposes. In one aspect, modifications of gene expressioncan be achieved by designing complementary sequences or antisensemoleculs (DNA, RNA, PNA, or modified oligonucleotides) to the coding orregulatory regions of the gene encoding TRICH. Such technology is wellknown in the art, and antisense oligonucleotides or larger fragments canbe designed from various locations along the coding or control regionsof sequences encoding TRICH. (See, e.g., Agrawal, S., ed. (1996)Antisense Therapeutics, Humana Press Inc., Totawa N.J.)

[0217] In therapeutic use, any gene delivery system suitable forintroduction of the antisense sequences into appropriate target cellscan be used. Antisense sequences can be delivered intracellularly in theform of an expression plasmid which, upon transcription, produces asequence complementary to at least a portion of the cellular sequenceencoding the target protein. (See, e.g., Slater, J. E. et al. (1998) J.Allergy Cli. Immunol. 102(3):469-475; and Scanlon, K. J. et al. (1995)9(13):1288-1296.) Antisense sequences can also be introducedintracellularly through the use of viral vectors, such as retrovirus andadeno-associated virus vectors. (See, e.g., Miller, A. D. (1990) Blood76:271; Ausubel, supra; Uckert, W. and W. Walther (1994) Pharmacol.Ther. 63(3):323-347.) Other gene delivery mechanisms includeliposome-derived systems, artificial viral envelopes, and other systemsknown in the art. (See, e.g., Rossi, J. J. (1995) Br. Med. Bull.51(1):217-225; Boado, R. J. et al. (1998) J. Pharm. Sci.87(11):1308-1315; and Morris, M. C. et al. (1997) Nucleic Acids Res.25(14):2730-2736.)

[0218] In another embodiment of the invention, polynucleotides encodingTRICH may be used for somatic or germline gene therapy. Gene therapy maybe performed to (i) correct a genetic deficiency (e.g., in the cases ofsevere combined immunodeficiency (SCID)-X1 disease characterized byX-linked inheritance (Cavazzana-Calvo, M. et al. (2000) Science288:669-672), severe combined immunodeficiency syndrome associated withan inherited adenosine deaminase (ADA) deficiency (Blaese, R. M. et al.(1995) Science 270:475480; Bordignon, C. et al. (1995) Science270:470475), cystic fibrosis (Zabner, J. et al. (1993) Cell 75:207-216;Crystal, R. G. et al. (1995) Hum. Gene Therapy 6:643-666; Crystal, R. G.et al. (1995) Hum. Gene Therapy 6:667-703), thalassamias, familialhypercholesterolemia, and hemophilia resulting from Factor VIII orFactor IX deficiencies (Crystal, R. G. (1995) Science 270:404410; Verma,I. M. and N. Somia (1997) Nature 389:239-242)), (ii) express aconditionally lethal gene product (e.g., in the case of cancers whichresult from unregulated cell proliferation), or (iii) express a proteinwhich affords protection against intracellular parasites (e.g., againsthuman retroviruses, such as human immunodeficiency virus (HIV)(Baltimore, D. (1988) Nature 335:395-396; Poeschla, E. et al. (1996)Proc. Natl. Acad. Sci. USA 93:11395-11399), hepatitis B or C virus (HBV,HCV); fungal parasites, such as Candida albicans and Paracoccidioidesbrasiliensis; and protozoan parasites such as

[0219] Plasmodium falciparum and Trypanosoma cruzi). In the case where agenetic deficiency in TRICH expression or regulation causes disease, theexpression of TRICH from an appropriate population of transduced cellsmay alleviate the clinical manifestations caused by the geneticdeficiency.

[0220] In a further embodiment of the invention, diseases or disorderscaused by deficiencies in TRICH are treated by constructing mammalianexpression vectors encoding TRICH and introducing these vectors bymechanical means into TRICH-deficient cells. Mechanical transfertechnologies for use with cells in vivo or ex vitro include (i) directDNA microinjection into individual cells, (ii) ballistic gold particledelivery, (iii) liposome-mediated transfection, (iv) receptor-mediatedgene transfer, and (v) the use of DNA transposons Morgan, R. A. and W.F. Anderson (1993) Annu. Rev. Biochem. 62:191-217; Ivics, Z. (1997) Cell91:501-510; Boulay, J-L. and H. Récipon (1998) Curr. Opin. Biotechnol.9:445450).

[0221] Expression vectors that may be effective for the expression ofTRICH include, but are not limited to, the PCDNA 3.1, EPITAG, PRCCMV2,PREP, PVAX vectors (Invitrogen, Carlsbad Calif.), PCMV-SCRIPT, PCMV-TAG,PEGSH/PERV (Stratagene, La Jolla Calif.), and PTET-OFF, PTET-ON, PTRE2,PTRE2-LUC, PTK-HYG (Clontech, Palo Alto Calif.). TRICH may be expressedusing (i) a constitutively active promoter, (e.g., from cytomegalovirus(CMV), Rous sarcoma virus (RSV), SV40 virus, thymidine kinase (TK), orβ-actin genes), (ii) an inducible promoter (e.g., thetetracycline-regulated promoter (Gossen, M. and H. Bujard (1992) Proc.Natl. Acad. Sci. U.S.A. 89:5547-5551; Gossen, M. et al. (1995) Science268:1766-1769; Rossi, F. M. V. and H. M. Blau (1998) Curr. Opin.Biotechnol. 9:451-456), commercially available in the T-REX plasmid(Invitrogen)); the ecdysone-inducible promoter (available in theplasmids PVGRXR and PIND; Invitrogen); the FK506/rapamycin induciblepromoter; or the RU486/mifepristone inducible promoter (Rossi, F. M. V.and Blau, H. M. supra)), or (iii) a tissue-specific promoter or thenative promoter of the endogenous gene encoding TRICH from a normalindividual.

[0222] Commercially available liposome transformation kits (e.g., thePERFECT LIPID TRANSFECTION KIT, available from Invitrogen) allow onewith ordinary skill in the art to deliver polynucleotides to targetcells in culture and require minimal effort to optimize experimentalparameters. In the alternative, transformation is performed using thecalcium phosphate method (Graham, F. L. and A. J. Eb (1973) Virology52:456-467), or by electroporation (Neumann, E. et al. (1982) EMBO J.1:841-845). The introduction of DNA to primary cells requiresmodification of these standardized mammalian transfection protocols.

[0223] In another embodiment of the invention, diseases or disorderscaused by genetic defects with respect to TRICH expression are treatedby constructing a retrovirus vector consisting of (i) the polynucleotideencoding TRICH under the control of an independent promoter or theretrovirus long terminal repeat (LTR) promoter, (ii) appropriate RNApackaging signals, and (iii) a Rev-responsive element (RRE) along withadditional retrovirus cis-acting RNA sequences and coding sequencesrequired for efficient vector propagation. Retrovirus vectors (e.g., PFBand PFBNEO) are commercially available (Stratagene) and are based onpublished data (Riviere, I. et al. (1995) Proc. Natl. Acad. Sci. U.S.A92:6733-6737), incorporated by reference herein. The vector ispropagated in an appropriate vector producing cell line (VPCL) thatexpresses an envelope gene with a tropism for receptors on the targetcells or a promiscuous envelope protein such as VSVg (Armentano, D. etal. (1987) J. Virol. 61:1647-1650; Bender, M. A. et al. (1987) J. Virol.61:1639-1646; Adam, M. A. and A. D. Miller (1988) J. Virol.62:3802-3806; Dull, T. et al. (1998) J. Virol. 72:8463-8471; Zufferey,R. et al. (1998) J. Virol. 72:9873-9880). U.S. Pat. No. 5,910,434 toRigg (“Method for obtaining retrovirus packaging cell lines producinghigh transducing efficiency retroviral supernatant”) discloses a methodfor obtaining retrovirus packaging cell lines and is hereby incorporatedby reference. Propagation of retrovirus vectors, transduction of apopulation of cells (e.g., CD4⁺ T-cells), and the return of transducedcells to a patient are procedures well known to persons skilled in theart of gene therapy and have been well documented (Ranga, U. et al.(1997) J. Virol. 71:7020-7029; Bauer, G. et al. (1997) Blood89:2259-2267; Bonyhadi, M. L. (1997) J. Virol. 71:4707-4716; Ranga, U.et al. (1998) Proc. Natl. Acad. Sci. U.S.A. 95:1201-1206; Su, L. (1997)Blood 89:2283-2290).

[0224] In the alternative, an adenovirus-based gene therapy deliverysystem is used to deliver polynucleotides encoding TRICH to cells whichhave one or more genetic abnormalities with respect to the expression ofTRICH. The construction and packaging of adenovirus-based vectors arewell known to those with ordinary skill in the art. Replicationdefective adenovirus vectors have proven to be versatile for importinggenes encoding immunoregulatory proteins into intact islets in thepancreas (Csete, M. E. et al. (1995) Transplantation 27:263-268).Potentially useful adenoviral vectors are described in U.S. Pat. No.5,707,618 to Armentano (“Adenovirus vectors for gene therapy”), herebyincorporated by reference. For adenoviral vectors, see also Antinozzi,P. A. et al. (1999) Annu. Rev. Nutr. 19:511-544 and Verma, I. M. and N.Somia (1997) Nature 18:389:239-242, both incorporated by referenceherein.

[0225] In another alternative, a herpes-based, gene therapy deliverysystem is used to deliver polynucleotides encoding TRICH to target cellswhich have one or more genetic abnormalities with respect to theexpression of TRICH. The use of herpes simplex virus (HSV)-based vectorsmay be especially valuable for introducing TRICH to cells of the centralnervous system, for which HSV has a tropism. The construction andpackaging of herpes-based vectors are well known to those with ordinaryskill in the art. A replication-competent herpes simplex virus (HSV)type 1-based vector has been used to deliver a reporter gene to the eyesof primates (Liu, X. et al. (1999) Exp. Eye Res. 169:385-395). Theconstruction of a HSV-1 virus vector has also been disclosed in detailin U.S. Pat. No. 5,804,413 to DeLuca (“Herpes simplex virus strains forgene transfer”), which is hereby incorporated by reference. U.S. Pat.No. 5,804,413 teaches the use of recombinant HSV d92 which consists of agenome containing at least one exogenous gene to be transferred to acell under the control of the appropriate promoter for purposesincluding human gene therapy. Also taught by this patent are theconstruction and use of recombinant HSV strains deleted for ICP4, ICP27and ICP22. For HSV vectors, see also Goins, W. F. et al. (1999) J.Virol. 73:519-532 and Xu, H. et al. (1994) Dev. Biol. 163:152-161,hereby incorporated by reference. The manipulation of cloned herpesvirussequences, the generation of recombinant virus following thetransfection of multiple plasmids containing different segments of thelarge herpesvirus genomes, the growth and propagation of herpesvirus,and the infection of cells with herpesvirus are techniques well known tothose of ordinary skill in the art.

[0226] In another alternative, an alphavirus (positive, single-strandedRNA virus) vector is used to deliver polynucleotides encoding TRICH totarget cells. The biology of the prototypic alphavirus, Semliki ForestVirus (SFV), has been studied extensively and gene transfer vectors havebeen based on the SFV genome (Garoff, H. and K. -J. Li (1998) Curr.Opin. Biotechnol. 9:464-469). During alphavirus RNA replication, asubgenomic RNA is generated that normally encodes the viral capsidproteins. This subgenomic RNA replicates to higher levels than the fulllength genomic RNA, resulting in the overproduction of capsid proteinsrelative to the viral proteins with enzymatic activity (e.g., proteaseand polymerase). Similarly, inserting the coding sequence for TRICH intothe alphavirus genome in place of the capsid-coding region results inthe production of a large number of TRICH-coding RNAs and the synthesisof high levels of TRICH in vector transduced cells. While alphavirusinfection is typically associated with cell lysis within a few days, theability to establish a persistent infection in hamster normal kidneycells (BHK-21) with a variant of Sindbis virus (SIN) indicates that thelytic replication of alphaviruses can be altered to suit the needs ofthe gene therapy application (Dryga, S. A. et al. (1997) Virology228:74-83). The wide host range of alphaviruses will allow theintroduction of TRICH into a variety of cell types. The specifictransduction of a subset of cells in a population may require thesorting of cells prior to transduction. The methods of manipulatinginfectious cDNA clones of alphaviruses, performing alphavirus cDNA andRNA transfections, and performing alphavirus infections, are well knownto those with ordinary skill in the art.

[0227] Oligonucleotides derived from the transcription initiation site,e.g., between about positions −10 and +10 from the start site, may alsobe employed to inhibit gene expression. Similarly, inhibition can beachieved using triple helix base-pairing methodology. Triple helixpairing is useful because it causes inhibition of the ability of thedouble helix to open sufficiently for the binding of polymerases,transcription factors, or regulatory molecules. Recent therapeuticadvances using triplex DNA have been described in the literature. (See,e.g., Gee, J. E. et al. (1994) in Huber, B. E. and B. I. Carr, Molecularand Immunologic Approaches, Futura Publishing, Mt. Kisco N.Y., pp.163-177.) A complementary sequence or antisense molecule may also bedesigned to block translation of mRNA by preventing the transcript frombinding to ribosomes.

[0228] Ribozymes, enzymatic RNA molecules, may also be used to catalyzethe specific cleavage of RNA. The mechanism of ribozyme action involvessequence-specific hybridization of the ribozyme molecule tocomplementary target RNA, followed by endonucleolytic cleavage. Forexample, engineered hammerhead motif ribozyme molecules may specificallyand efficiently catalyze endonucleolytic cleavage of sequences encodingTRICH.

[0229] Specific ribozyme cleavage sites within any potential RNA targetare initially identified by scanning the target molecule for ribozymecleavage sites, including the following sequences: GUA, GUU, and GUC.Once identified, short RNA sequences of between 15 and 20ribonucleotides, corresponding to the region of the target genecontaining the cleavage site, may be evaluated for secondary structuralfeatures which may render the oligonucleotide inoperable. Thesuitability of candidate targets may also be evaluated by testingaccessibility to hybridization with complementary oligonucleotides usingribonuclease protection assays.

[0230] Complementary ribonucleic acid molecules and ribozymes of theinvention may be prepared by any method known in the art for thesynthesis of nucleic acid molecules. These include techniques forchemically synthesizing oligonucleotides such as solid phasephosphoramidite chemical synthesis. Alternatively, RNA molecules may begenerated by in vitro and in vivo transcription of DNA sequencesencoding TRICH. Such DNA sequences may be incorporated into a widevariety of vectors with suitable RNA polymerase promoters such as 1 orSP6. Alternatively, these cDNA constructs that synthesize complementaryRNA, constitutively or inducibly, can be introduced into cell lines,cells, or tissues.

[0231] RNA molecules may be modified to increase intracellular stabilityand half-life. Possible modifications include, but are not limited to,the addition of flanking sequences at the 5′ and/or 3′ ends of themolecule, or the use of phosphorothioate or 2′O-methyl rather thanphosphodiesterase linkages within the backbone of the molecule. Thisconcept is inherent in the production of PNAs and can be extended in anof these molecules by the inclusion of nontraditional bases such asinosine, queosine, and wybutosine, as well as acetyl-, methyl-, thio-,and similarly modified forms of adenine, cytidine, guanine, thymine, anduridine which are not as easily recognized by endogenous endonucleases.

[0232] An additional embodiment of the invention encompasses a methodfor screening for a compound which is effective in altering expressionof a polynucleotide encoding TRICH. Compounds which may be effective inaltering expression of a specific polynucleotide may include, but arenot limited to, oligonucleotides, antisense oligonucleotides, triplehelix-forming oligonucleotides, transcription factors and otherpolypeptide transcriptional regulators, and non-macromolecular chemicalentities which are capable of interacting with specific polynucleotidesequences. Effective compounds may alter polynucleotide expression byacting as either inhibitors or promoters of polynucleotide expression.Thus, in the treatment of disorders associated with increased TRICHexpression or activity, a compound which specifically inhibitsexpression of the polynucleotide encoding TRICH may be therapeuticallyuseful, and in the treament of disorders associated with decreased TRICHexpression or activity, a compound which specifically promotesexpression of the polynucleotide encoding TRICH may be therapeuticallyuseful.

[0233] At least one, and up to a plurality, of test compounds may bescreened for effectiveness in altering expression of a specificpolynucleotide. A test compound may be obtained by any method commonlyknown in the art, including chemical modification of a compound known tobe effective in altering polynucleotide expression; selection from anexisting, commercially-available or proprietary library ofnaturally-occurring or non-natural chemical compounds; rational designof a compound based on chemical and/or structural properties of thetarget polynucleotide; and selection from a library of chemicalcompounds created combinatorially or randomly. A sample comprising apolynucleotide encoding TRICH is exposed to at least one test compoundthus obtained. The sample may comprise, for example, an intact orpermeabilized cell, or an in vitro cell-free or reconstitutedbiochemical system. Alterations in the expression of a polynucleotideencoding TRICH are assayed by any method commonly known in the art.Typically, the expression of a specific nucleotide is detected byhybridization with a probe having a nucleotide sequence complementary tothe sequence of the polynucleotide encoding TRICH. The amount ofhybridization may be quantified, thus forming the basis for a comparisonof the expression of the polynucleotide both with and without exposureto one or more test compounds. Detection of a change in the expressionof a polynucleotide exposed to a test compound indicates that the testcompound is effective in altering the expression of the polynucleotide.A screen for a compound effective in altering expression of a specificpolynucleotide can be carried out, for example, using aSchizosaccharomvces pombe gene expression system (Atkins, D. et al.(1999) U.S. Pat. No. 5,932,435; Arndt, G. M. et al. (2000) Nucleic AcidsRes. 28:E15) or a human cell line such as HeLa cell (Clarke, M. L. etal. (2000) Biochem. Biophys. Res. Commun. 268:8-13). A particularembodiment of the present invention involves screening a combinatoriallibrary of oligonucleotides (such as deoxyribonucleotides,ribonucleotides, peptide nucleic acids, and modified oligonucleotides)for antisense activity against a specific polynucleotide sequence(Bruice, T. W. et al. (1997) U.S. Pat. No. 5,686,242; Bruice, T. W. etal. (2000) U.S. Pat. No. 6,022,691).

[0234] Many methods for introducing vectors into cells or tissues areavailable and equally suitable for use in vivo, in vitro, and ex vivo.For ex vivo therapy, vectors may be introduced into stem cells takenfrom the patient and clonally propagated for autologous transplant backinto that same patient. Delivery by transfection, by liposomeinjections, or by polycationic amino polymers may be achieved usingmethods which are well known in the art. (See, e.g., Goldman, C. K. etal. (1997) Nat. Biotechnol. 15:462-466.)

[0235] Any of the therapeutic methods described above may be applied toany subject in need of such therapy, including, for example, mammalssuch as humans, dogs, cats, cows, horses, rabbits, and monkeys.

[0236] An additional embodiment of the invention relates to theadministration of a composition which generally comprises an activeingredient formulated with a pharmaceutically acceptable excipient.Excipients may include, for example, sugars, starches, celluloses, gums,and proteins. Various formulations are commonly known and are thoroughlydiscussed in the latest edition of Remington's Pharmaceutical Sciences(Maack Publishing, Easton Pa.). Such compositions may consist of TRICH,antibodies to TRICH, and mimetics, agonists, antagonists, or inhibitorsof TRICH.

[0237] The compositions utilized in this invention may be administeredby any number of routes including, but not limited to, oral,intravenous, intramuscular, intra-arterial, intramedullary, intrathecal,intraventricular, pulmonary, transdermal, subcutaneous, intraperitoneal,intranasal, enteral, topical, sublingual, or rectal means.

[0238] Compositions for pulmonary administration may be prepared inliquid or dry powder form. These compositions are generally aerosolizedimmediately prior to inhalation by the patient. In the case of smallmolecules (e.g. traditional low molecular weight organic drugs), aerosoldelivery of fast-acting formulations is well-known in the art. In thecase of macromolecules (e.g. larger peptides and proteins), recentdevelopments in the field of pulmonary delivery via the alveolar regionof the lung have enabled the practical delivery of drugs such as insulinto blood circulation (see, e.g., Patton. J. S. et al., U.S. Pat. No.5,997,848). Pulmonary delivery has the advantage of administrationwithout needle injection, and obviates the need for potentially toxicpenetration enhancers.

[0239] Compositions suitable for use in the invention includecompositions wherein the active ingredients are contained in aneffective amount to achieve the intended purpose. The determination ofan effective dose is well within the capability of those skilled in theart.

[0240] Specialized forms of compositions may be prepared for directintracellular delivery of macromolecules comprising TRICH or fragmentsthereof. For example, liposome preparations containing acell-impermeable macromolecule may promote cell fusion and intracellulardelivery of the macromolecule. Alternatively, TRICH or a fragmentthereof may be joined to a short cationic N-terminal portion from theHIV Tat-1 protein. Fusion proteins thus generated have been found totransduce into the cells of all tissues, including the brain, in a mousemodel system (Schwarze, S. R. et al. (1999) Science 285:1569-1572).

[0241] For any compound, the therapeutically effective dose can beestimated initially either in cell culture assays, e.g., of neoplasticcells, or in animal models such as mice, rats, rabbits, dogs, monkeys,or pigs. An animal model may also be used to determine the appropriateconcentration range and route of administration. Such information canthen be used to determine useful doses and routes for administration inhumans.

[0242] A therapeutically effective dose refers to that amount of activeingredient, for example TRICH or fragments thereof, antibodies of TRICH,and agonists, antagonists or inhibitors of TRICH, which ameliorates thesymptoms or condition. Therapeutic efficacy and toxicity may bedetermined by standard pharmaceutical procedures in cell cultures orwith experimental animals, such as by calculating the ED₅₀ (the dosetherapeutically effective in 50% of the population) or LD₅₀ (the doselethal to 50% of the population) statistics. The dose ratio of toxic totherapeutic effects is the therapeutic index, which can be expressed asthe LD₅₀/ED₅₀ ratio. Compositions which exhibit large therapeuticindices are preferred. The data obtained from cell culture assays andanimal studies are used to formulate a range of dosage for human use.The dosage contained in such compositions is preferably within a rangeof circulating concentrations that includes the ED₅₀ with lithe or notoxicity. The dosage varies within this range depending upon the dosageform employed, the sensitivity of the patient, and the route ofadministration.

[0243] The exact dosage will be determined by the practitioner, in lightof factors related to the subject requiring treatment. Dosage andadministration are adjusted to provide sufficient levels of the activemoiety or to maintain the desired effect. Factors which may be takeninto account include the severity of the disease state, the generalhealth of the subject, the age, weight, and gender of the subject, timeand frequency of administration, drug combination(s), reactionsensitivities, and response to therapy. Long-acting compositions may beadministered every 3 to 4 days, every week, or biweekly depending on thehalf-life and clearance rate of the particular formulation.

[0244] Normal dosage amounts may vary from about 0.1 μg to 100,000 μg,up to a total dose of about 1 gram, depending upon the route ofadministration. Guidance as to particular dosages and methods ofdelivery is provided in the literature and generally available topractitioners in the art. Those skilled in the art will employ differentformulations for nucleotides than for proteins or their inhibitors.Similarly, delivery of polynucleotides or polypeptides will be specificto particular cells, conditions, locations, etc.

[0245] Diagnostics

[0246] In another embodiment, antibodies which specifically bind TRICHmay be used for the diagnosis of disorders characterized by expressionof TRICH, or in assays to monitor patients being treated with TRICH oragonists, antagonists, or inhibitors of TRICH. Antibodies useful fordiagnostic purposes may be prepared in the same manner as describedabove for therapeutics. Diagnostic assays for TRICH include methodswhich utilize the antibody and a label to detect TRICH in human bodyfluids or in extracts of cells or tissues. The antibodies may be usedwith or without modification, and may be labeled by covalent ornon-covalent attachment of a reporter molecule. A wide variety ofreporter molecules, several of which are described above, are known inthe art and may be used.

[0247] A variety of protocols for measuring TRICH, including ELISAs,RIAs, and FACS, are known in the art and provide a basis for diagnosingaltered or abnormal levels of TRICH expression. Normal or standardvalues for TRICH expression are established by combining body fluids orcell extracts taken from normal mammalian subjects, for example, humansubjects, with antibodies to TRICH under conditions suitable for complexformation. The amount of standard complex formation may be quantitatedby various methods, such as photometric means. Quantities of TRICHexpressed in subject, control, and disease samples from biopsied tissuesare compared with the standard values. Deviation between standard andsubject values establishes the parameters for diagnosing disease.

[0248] In another embodiment of the invention, the polynucleotidesencoding TRICH may be used for diagnostic purposes. The polynucleotideswhich may be used include oligonucleotide sequences, complementary RNAand DNA molecules, and PNAs. The polynucleotides may be used to detectand quantify gene expression in biopsied tissues in which expression ofTRICH may be correlated with disease. The diagnostic assay may be usedto determine absence, presence, and excess expression of TRICH, and tomonitor regulation of TRICH levels during therapeutic intervention.

[0249] In one aspect, hybridization with PCR probes which are capable ofdetecting polynucleotide sequences, including genomic sequences,encoding TRICH or closely related molecules may be used to identifynucleic acid sequences which encode TRICH. The specificity of the probe,whether it is made from a highly specific region, e.g., the 5′regulatory region, or from a less specific region, e.g., a conservedmotif, and the stringency of the hybridization or amplification willdetermine whether the probe identifies only naturally occurringsequences encoding TRICH, allelic variants, or related sequences.

[0250] Probes may also be used for the detection of related sequences,and may have at least 50% sequence identity to any of the TRICH encodingsequences. The hybridization probes of the subject invention may be DNAor RNA and may be derived from the sequence of SEQ ID NO:28-54 or fromgenomic sequences including promoters, enhancers, and introns of theTRICH gene.

[0251] Means for producing specific hybridization probes for DNAsencoding TRICH include the cloning of polynucleotide sequences encodingTRICH or TRICH derivatives into vectors for the production of mRNAprobes. Such vectors are known in the art, are commercially available,and may be used to synthesize RNA probes in vitro by means of theaddition of the appropriate RNA polymerases and the appropriate labelednucleotides. Hybridization probes may be labeled by a variety ofreporter groups, for example, by radionuclides such as ³²P or ³⁵S, or byenzymatic labels, such as alkaline phosphatase coupled to the probe viaavidin/biotin coupling systems, and the like.

[0252] Polynucleotide sequences encoding TRICH may be used for thediagnosis of disorders associated with expression of TRICH. Examples ofsuch disorders include, but are not limited to, a transport disordersuch as akinesia, amyotrophic lateral sclerosis, ataxia telangiectasia,cystic fibrosis, Becker's muscular dystrophy, Bell's palsy,Charcot-Marie Tooth disease, diabetes mellitus, diabetes insipidus,diabetic neuropathy, Duchenne muscular dystrophy, hyperkalemic periodicparalysis, normokalemic periodic paralysis, Parkinson's disease,malignant hyperthermia, multidrug resistance, myasthenia gravis,myotonic dystrophy, catatonia, tardive dyskinesia, dystonias, peripheralneuropathy, cerebral neoplasms, prostate cancer, cardiac disordersassociated with transport, e.g., angina, bradyarrythmia, tachyarrythmia,hypertension, Long QT syndrome, myocarditis, cardiomyopathy, nemalinemyopathy, centronuclear myopathy, lipid myopathy, mitochondrialmyopathy, thyrotoxic myopathy, ethanol myopathy, dermatomyositis,inclusion body myositis, infectious myositis, polymyositis, neurologicaldisorders associated with transport, e.g., Alzheimer's disease, amnesia,bipolar disorder, dementia, depression, epilepsy, Tourette's disorder,paranoid psychoses, and schizophrenia, and other disorders associatedwith transport, e.g., neurofibromatosis, postherpetic neuralgia,trigeminal neuropathy, sarcoidosis, sickle cell anemia, Wilson'sdisease, cataracts, infertility, pulmonary artery stenosis,sensorineural autosomal deafness, hyperglycemia, hypoglycemia, Grave'sdisease, goiter, Cushing's disease, Addison's disease, glucose-galactosemalabsorption syndrome, hypercholesterolemia, adrenoleukodystrophy,Zellweger syndrome, Menkes disease, occipital horn syndrome, von Gierkedisease, cystinuria, iminoglycinuria, Hartup disease, and Fanconidisease; a neurological disorder such as epilepsy, ischemiccerebrovascular disease, stroke, cerebral neoplasms, Alzheimer'sdisease, Pick's disease, Huntington's disease, dementia, Parkinson'sdisease and other extrapyramidal disorders, amyotrophic lateralsclerosis and other motor neuron disorders, progressive neural muscularatrophy, retinitis pigmentosa, hereditary ataxias, multiple sclerosisand other demyelinating diseases, bacterial and viral meningitis, brainabscess, subdural empyema, epidural abscess, suppurative intracranialthrombophlebitis, myelitis and radiculitis, viral central nervous systemdisease, prion diseases including kuru, Creutzfeldt-Jakob disease, andGerstmann-Straussler-Scheinker syndrome, fatal familial insomnia,nutritional and metabolic diseases of the nervous system,neurofibromatosis, tuberous sclerosis, cerebelloretinalhemangioblastomatosis, encephalotrigeminal syndrome, mental retardationand other developmental disorders of the central nervous systemincluding Down syndrome, cerebral palsy, neuroskeletal disorders,autonomic nervous system disorders, cranial nerve disorders, spinal corddiseases, muscular dystrophy and other neuromuscular disorders,peripheral nervous system disorders, dermatomyositis and polymyositis,inherited, metabolic, endocrine, and toxic myopathies, myastheniagravis, periodic paralysis, mental disorders including mood, anxiety,and schizophrenic disorders, seasonal affective disorder (SAD),akathesia, amnesia, catatonia, diabetic neuropathy, tardive dyskinesia,dystonias, paranoid psychoses, postherpetic neuralgia, Tourette'sdisorder, progressive supranuclear palsy, corticobasal degeneration, andfamilial frontotemporal dementia; a muscle disorder such ascardiomyopathy, myocarditis, Duchenne's muscular dystrophy, Becker'smuscular dystrophy, myotonic dystrophy, central core disease, nemalinemyopathy, centronuclear myopathy, lipid myopathy, mitochondrialmyopathy, infectious myositis, polymyositis, dermatomyositis, inclusionbody myositis, thyrotoxic myopathy, ethanol myopathy, angina,anaphylactic shock, arrhythmias, asthma, cardiovascular shock, Cushing'ssyndrome, hypertension, hypoglycemia, myocardial infarction, migraine,pheochromocytoma, and myopathies including encephalopathy, epilepsy,Kearns-Sayre syndrome, lactic acidosis, myoclonic disorder,ophthalmoplegia, and acid maltase deficiency (AMD, also known as Pompe'sdisease); and an immunological disorder such as acquiredimmunodeficiency syndrome (AIDS), Addison's disease, adult respiratorydistress syndrome, allergies, ankylosing spondylitis, amyloidosis,anemia, asthma, atherosclerosis, autoimmune hemolytic anemia, autoimmunethyroiditis, autoimmune polyendocrinopathy-candidiasis-ectodermaldystrophy (APECED), bronchitis, cholecystitis, contact dermatitis,Crohn's disease, atopic dermatitis, dermatomyositis, diabetes mellitus,emphysema, episodic lymphopenia with lymphocytotoxins, erythroblastosisfetalis, erythema nodosum, atrophic gastritis, glomerulonephritis,Goodpasture's syndrome, gout, Graves' disease, Hashimoto's thyroiditis,hypereosinophilia, irritable bowel syndrome, multiple sclerosis,myasthenia gravis, myocardial or pericardial inflammation,osteoarthritis, osteoporosis, pancreatitis, polymyositis, psoriasis,Reiter's syndrome, rheumatoid arthritis, scleroderma, Sjögren'ssyndrome, systemic anaphylaxis, systemic lupus erythematosus, systemicsclerosis, thrombocytopenic purpura, ulcerative colitis, uveitis, Wernersyndrome, complications of cancer, hemodialysis, and extracorporealcirculation, viral, bacterial, fungal, parasitic, protozoal, andhelminthic infections, and trauma. The polynucleotide sequences encodingTRICH may be used in Southern or northern analysis, dot blot, or othermembrane-based technologies; in PCR technologies; in dipstick, pin, andmultiformat ELISA-like assays; and in microarrays utilizing fluids ortissues from patients to detect altered TRICH expression. Suchqualitative or quantitative methods are well known in the art.

[0253] In a particular aspect, the nucleotide sequences encoding TRICHmay be useful in assays that detect the presence of associateddisorders, particularly those mentioned above. The nucleotide sequencesencoding TRICH may be labeled by standard methods and added to a fluidor tissue sample from a patient under conditions suitable for theformation of hybridization complexes. After a suitable incubationperiod, the sample is washed and the signal is quantified and comparedwith a standard value. If the amount of signal in the patient sample issignificantly altered in comparison to a control sample then thepresence of altered levels of nucleotide sequences encoding TRICH in thesample indicates the presence of the associated disorder. Such assaysmay also be used to evaluate the efficacy of a particular therapeutictreatment regimen in animal studies, in clinical trials, or to monitorthe treatment of an individual patient.

[0254] In order to provide a basis for the diagnosis of a disorderassociated with expression of TRICH, a normal or standard profile forexpression is established. This may be accomplished by combining bodyfluids or cell extracts taken from normal subjects, either animal orhuman, with a sequence, or a fragment thereof, encoding TRICH, underconditions suitable for hybridization or amplification. Standardhybridization may be quantified by comparing the values obtained fromnormal subjects with values from an experiment in which a known amountof a substantially purified polynucleotide is used. Standard valuesobtained in this manner may be compared with values obtained fromsamples from patients who are symptomatic for a disorder. Deviation fromstandard values is used to establish the presence of a disorder.

[0255] Once the presence of a disorder is established and a treatmentprotocol is initiated, hybridization assays may be repeated on a regularbasis to determine if the level of expression in the patient begins toapproximate that which is observed in the normal subject. The resultsobtained from successive assays may be used to show the efficacy oftreatment over a period ranging from several days to months.

[0256] With respect to cancer, the presence of an abnormal amount oftranscript (either under- or overexpressed) in biopsied tissue from anindividual may indicate a predisposition for the development of thedisease, or may provide a means for detecting the disease prior to theappearance of actual clinical symptoms. A more definitive diagnosis ofthis type may allow health professionals to employ preventative measuresor aggressive treatment earlier thereby preventing the development orfurther progression of the cancer.

[0257] Additional diagnostic uses for oligonucleotides designed from thesequences encoding TRICH may involve the use of PCR. These oligomers maybe chemically synthesized, generated enzymatically, or produced invitro. Oligomers will preferably contain a fragment of a polynucleotideencoding TRICH, or a fragment of a polynucleotide complementary to thepolynucleotide encoding TRICH, and will be employed under optimizedconditions for identification of a specific gene or condition. Oligomersmay also be employed under less stringent conditions for detection orquantification of closely related DNA or RNA sequences.

[0258] In a particular aspect, oligonucleotide primers derived from thepolynucleotide sequences encoding TRICH may be used to detect singlenucleotide polymorphisms (SNPs). SNPs are substitutions, insertions anddeletions that are a frequent cause of inherited or acquired geneticdisease in humans. Methods of SNP detection include, but are not limitedto, single-stranded conformation polymorphism (SSCP) and fluorescentSSCP (FSSCP) methods. In SSCP, oligonucleotide primers derived from thepolynucleotide sequences encoding TRICH are used to amplify DNA usingthe polymerase chain reaction (PCR). The DNA may be derived, forexample, from diseased or normal tissue, biopsy samples, bodily fluids,and the like. SNPs in the DNA cause differences in the secondary andtertiary structures of PCR products in single-stranded form, and thesedifferences are detectable using gel electrophoresis in non-denaturinggels. In fSCCP, the oligonucleotide priers are fluorescently labeled,which allows detection of the amplimers in high-throughput equipmentsuch as DNA sequencing machines. Additionally, sequence databaseanalysis methods, termed in silico SNP (isSNP), are capable ofidentifying polymorphisms by comparing the sequence of individualoverlapping DNA fragments which assemble into a common consensussequence. These computer-based methods filter out sequence variationsdue to laboratory preparation of DNA and sequencing errors usingstatistical models and automated analyses of DNA sequence chromatograms.In the alternative, SNPs may be detected and characterized by massspectrometry using, for example, the high throughput MASSARRAY system(Sequenom, Inc., San Diego Calif.).

[0259] Methods which may also be used to quantify the expression ofTRICH include radiolabeling or biotinylating nucleotides,coamplification of a control nucleic acid, and interpolating resultsfrom standard curves. (See, e.g., Melby, P. C. et al. (1993) J. Immunol.Methods 159:235-244; Duplaa, C. et al. (1993) Anal. Biochem.212:229-236.) The speed of quantitation of multiple samples may beaccelerated by running the assay in a high-throughput format where theoligomer or polynucleotide of interest is presented in various dilutionsand a spectrophotometric or colorimetric response gives rapidquantitation.

[0260] In further embodiments, oligonucleotides or longer fragmentsderived from any of the polynucleotide sequences described herein may beused as elements on a microarray. The microarray can be used intranscript imaging techniques which monitor the relative expressionlevels of large numbers of genes simultaneously as described below. Themicroarray may also be used to identify genetic variants, mutations, andpolymorphisms. This information may be used to determine gene function,to understand the genetic basis of a disorder, to diagnose a disorder,to monitor progression/regression of disease as a function of geneexpression, and to develop and monitor the activities of therapeuticagents in the treatment of disease. In particular, this information maybe used to develop a pharmacogenomic profile of a patient in order toselect the most appropriate and effective treatment regimen for thatpatient. For example, therapeutic agents which are highly effective anddisplay the fewest side effects may be selected for a patient based onhis/her pharmacogenomic profile.

[0261] In another embodiment, TRICH, fragments of TRICH, or antibodiesspecific for TRICH may be used as elements on a microarray. Themicroarray may be used to monitor or measure protein-proteininteractions, drug-target interactions, and gene expression profiles, asdescribed above.

[0262] A particular embodiment relates to the use of the polynucleotidesof the present invention to generate a transcript image of a tissue orcell type. A transcript image represents the global pattern of geneexpression by a particular tissue or cell type. Global gene expressionpatterns are analyzed by quantifying the number of expressed genes andtheir relative abundance under given conditions and at a given time.(See Seilhamer et al., “Comparative Gene Transcript Analysis,” U.S. Pat.No. 5,840,484, expressly incorporated by reference herein.) Thus atranscript image may be generated by hybridizing the polynucleotides ofthe present invention or their complements to the totality oftranscripts or reverse transcripts of a particular tissue or cell type.In one embodiment, the hybridization takes place in high-throughputformat, wherein the polynucleotides of the present invention or theircomplements comprise a subset of a plurality of elements on amicroarray. The resultant transcript image would provide a profile ofgene activity.

[0263] Transcript images may be generated using transcripts isolatedfrom tissues, cell lines, biopsies, or other biological samples. Thetranscript image may thus reflect gene expression in vivo, as in thecase of a tissue or biopsy sample, or in vitro, as in the case of a cellline.

[0264] Transcript images which profile the expression of thepolynucleotides of the present invention may also be used in conjunctionwith in vitro model systems and preclinical evaluation ofpharmaceuticals, as well as toxicological testing of industrial andnaturally-occurring environmental compounds. All compounds inducecharacteristic gene expression patterns, frequently termed molecularfingerprints or toxicant signatures, which are indicative of mechanismsof action and toxicity (Nuwaysir, E. F. et al. (1999) Mol. Carcinog.24:153-159; Steiner, S. and N. L. Anderson (2000) Toxicol. Lett.112-113:467-471, expressly incorporated by reference herein). If a testcompound has a signature similar to that of a compound with knowntoxicity, it is likely to share those toxic properties. Thesefingerprints or signatures are most useful and refined when they containexpression information from a large number of genes and gene families.Ideally, a genome-wide measurement of expression provides the highestquality signature. Even genes whose expression is not altered by anytested compounds are important as well, as the levels of expression ofthese genes are used to normalize the rest of the expression data. Thenormalization procedure is useful for comparison of expression dataafter treatment with different compounds. While the assignment of genefunction to elements of a toxicant signature aids in interpretation oftoxicity mechanisms, knowledge of gene function is not necessary for thestatistical matching of signatures which leads to prediction oftoxicity. (See, for example, Press Release 00-02 from the NationalInstitute of Environmental Health Sciences, released Feb. 29, 2000,available at http://www.niehs.nihgov/oc/news/toxchip.htm) Therefore, itis important and desirable in toxicological screening using toxicantsignatures to include all expressed gene sequences.

[0265] In one embodiment, the toxicity of a test compound is assessed bytreating a biological sample containing nucleic acids with the testcompound Nucleic acids that are expressed in the treated biologicalsample are hybridized with one or more probes specific to thepolynucleotides of the present invention, so that transcript levelscorresponding to the polynucleotides of the present invention may bequantified. The transcript levels in the treated biological sample arecompared with levels in an untreated biological sample. Differences inthe transcript levels between the two samples are indicative of a toxicresponse caused by the test compound in the treated sample.

[0266] Another particular embodiment relates to the use of thepolypeptide sequences of the present invention to analyze the proteomeof a tissue or cell type. The term proteome refers to the global patternof protein expression in a particular tissue or cell type. Each proteincomponent of a proteome can be subjected individually to furtheranalysis. Proteome expression patterns, or profiles, are analyzed byquantifying the number of expressed proteins and their relativeabundance under given conditions and at a given time. A profile of acell's proteome may thus be generated by separating and analyzing thepolypeptides of a particular tissue or cell type. In one embodiment, theseparation is achieved using two-dimensional gel electrophoresis, inwhich proteins from a sample are separated by isoelectric focusing inthe first dimension, and then according to molecular weight by sodiumdodecyl sulfate slab gel electrophoresis in the second dimension(Steiner and Anderson, supra). The proteins are visualized in the gel asdiscrete and uniquely positioned spots, typically by staining the gelwith an agent such as Coomassie Blue or silver or fluorescent stains.The optical density of each protein spot is generally proportional tothe level of the protein in the sample. The optical densities ofequivalently positioned protein spots from different samples, forexample, from biological samples either treated or untreated with a testcompound or therapeutic agent, are compared to identify any changes inprotein spot density related to the treatment. The proteins in the spotsare partially sequenced using, for example, standard methods employingchemical or enzymatic cleavage followed by mass spectrometry. Theidentity of the protein in a spot may be determined by comparing itspartial sequence, preferably of at least 5 contiguous amino acidresidues, to the polypeptide sequences of the present invention. In somecases, further sequence data may be obtained for definitive proteinidentification.

[0267] A proteomic profile may also be generated using antibodiesspecific for TRICH to quantify the levels of TRICH expression. In oneembodiment, the antibodies are used as elements on a microarray, andprotein expression levels are quantified by exposing the microarray tothe sample and detecting the levels of protein bound to each arrayelement (Lueking, A. et al. (1999) Anal. Biochem. 270:103-111; Mendoze,L. G. et al. (1999) Biotechniques 27:778-788). Detection may beperformed by a variety of methods known in the art, for example, byreacting the proteins in the sample with a thiol- or amino-reactivefluorescent compound and detecting the amount of fluorescence bound ateach array element.

[0268] Toxicant signatures at the proteome level are also useful fortoxicological screening, and should be analyzed in parallel withtoxicant signatures at the transcript level. There is a poor correlationbetween transcript and protein abundances for some proteins in sometissues (Anderson, N. L. and J. Seilhamer (1997) Electrophoresis18:533-537), so proteome toxicant signatures may be useful in theanalysis of compounds which do not significantly affect the transcriptimage, but which alter the proteomic profile. In addition, the analysisof transcripts in body fluids is difficult, due to rapid degradation ofmRNA, so proteomic profiling may be more reliable and informative insuch cases.

[0269] In another embodiment, the toxicity of a test compound isassessed by treating a biological sample containing proteins with thetest compound. Proteins that are expressed in the treated biologicalsample are separated so that the amount of each protein can bequantified. The amount of each protein is compared to the amount of thecorresponding protein in an untreated biological sample. A difference inthe amount of protein between the two samples is indicative of a toxicresponse to the test compound in the treated sample. Individual proteinsare identified by sequencing the amino acid residues of the individualproteins and comparing these partial sequences to the polypeptides ofthe present invention.

[0270] In another embodiment, the toxicity of a test compound isassessed by treating a biological sample containing proteins with thetest compound. Proteins from the biological sample are incubated withantibodies specific to the polypeptides of the present invention. Theamount of protein recognized by the antibodies is quantified. The amountof protein in the treated biological sample is compared with the amountin an untreated biological sample. A difference in the amount of proteinbetween the two samples is indicative of a toxic response to the testcompound in the treated sample.

[0271] Microarrays may be prepared, used, and analyzed using methodsknown in the art (See, e.g., Brennan, T. M. et al. (1995) U.S. Pat. No.5,474,796; Schena, M. et al. (1996) Proc. Natl. Acad. Sci. USA93:10614-10619; Baldeschweiler et al. (1995) PCT applicationWO95/251116; Shalon, D. et al. (1995) PCT application WO95/35505;Heller, R. A. et al. (1997) Proc. Natl. Acad. Sci. USA 94:2150-2155; andHeller, M. J. et al. (1997) U.S. Pat. No. 5,605,662.) Various types ofmicroarrays are well known and thoroughly described in DNA Microarrays:A Practical Approach, M. Schena, ed. (1999) Oxford University Press,London, hereby expressly incorporated by reference.

[0272] In another embodiment of the invention, nucleic acid sequencesencoding TRICH may be used to generate hybridization probes useful inmapping the naturally occurring genomic sequence. Either coding ornoncoding sequences may be used, and in some instances, noncodingsequences may be preferable over coding sequences. For example,conservation of a coding sequence among members of a multi-gene familymay potentially cause undesired cross hybridization during chromosomalmapping. The sequences may be mapped to a particular chromosome, to aspecific region of a chromosome, or to artificial chromosomeconstructions, e.g., human artificial chromosomes (HACs), yeastartificial chromosomes (YACs), bacterial artificial chromosomes (BACs),bacterial P1 constructions, or single chromosome cDNA libraries. (See,e.g., Harrington, J. J. et al. (1997) Nat. Genet. 15:345-355; Price, C.M. (1993) Blood Rev. 7:127-134; and Trask, B. J. (1991) Trends Genet7:149-154.) Once mapped, the nucleic acid sequences of the invention maybe used to develop genetic linkage maps, for example, which correlatethe inheritance of a disease state with the inheritance of a particularchromosome region or restriction fragment length polymorphism (RFLP).(See, for example, Lander, E. S. and D. Botstein (1986) Proc. Natl.Acad. Sci. USA 83:7353-7357.)

[0273] Fluorescent in situ hybridization (FISH) may be correlated withother physical and genetic map data. (See, e.g., Heinz-Ulrich, et al.(1995) in Meyers, supra, pp. 965-968.) Examples of genetic map data canbe found in various scientific journals or at the Online MendelianInheritance in Man (OMIM) World Wide Web site. Correlation between thelocation of the gene encoding TRICH on a physical map and a specificdisorder, or a predisposition to a specific disorder, may help definethe region of DNA associated with that disorder and thus may furtherpositional cloning efforts.

[0274] In situ hybridization of chromosomal preparations and physicalmapping techniques, such as linkage analysis using establishedchromosomal markers, may be used for extending genetic maps. Often theplacement of a gene on the chromosome of another mammalian species, suchas mouse, may reveal associated markers even if the exact chromosomallocus is not known. This information is valuable to investigatorssearching for disease genes using positional cloning or other genediscovery techniques. Once the gene or genes responsible for a diseaseor syndrome have been crudely localized by genetic linkage to aparticular genomic region, e.g., ataxia-telangiectasia to 11q22-23, anysequences mapping to that area may represent associated or regulatorygenes for further investigation (See, e.g., Gatti, R. A. et al. (1988)Nature 336:577-580.) The nucleotide sequence of the instant inventionmay also be used to detect differences in the chromosomal location dueto translocation, inversion, etc., among normal, carrier, or affectedindividuals.

[0275] In another embodiment of the invention, TRICH, its catalytic orimmunogenic fragments, or oligopeptides thereof can be used forscreening libraries of compounds in any of a variety of drug screeningtechniques. The fragment employed in such screening may be free insolution, affixed to a solid support, borne on a cell surface, orlocated intracellularly. The formation of binding complexes betweenTRICH and the agent being tested may be measured.

[0276] Another technique for drug screening provides for high throughputscreening of compounds having suitable binding affinity to the proteinof interest. (See, e.g., Geysen, et al. (1984) PCT applicationWO84/03564.) In this method, large numbers of different small testcompounds are synthesized on a solid substrate. The test compounds arereacted with TRICH, or fragments thereof, and washed. Bound TRICH isthen detected by methods well known in the art. Purified TRICH can alsobe coated directly onto plates for use in the aforementioned drugscreening techniques. Alternatively, non-neutralizing antibodies can beused to capture the peptide and immobilize it on a solid support.

[0277] In another embodiment, one may use competitive drug screeningassays in which neutralizing antibodies capable of binding TRICHspecifically compete with a test compound for binding TRICH. In thismanner, antibodies can be used to detect the presence of any peptidewhich shares one or more antigenic determinants with TRICH.

[0278] In additional embodiments, the nucleotide sequences which encodeTRICH may be used in any molecular biology techniques that have yet tobe developed, provided the new techniques rely on properties ofnucleotide sequences that are currently known, including, but notlimited to, such properties as the triplet genetic code and specificbase pair interactions.

[0279] Without further elaboration, it is believed that one skilled inthe art can, using the preceding description, utilize the presentinvention to its fullest extent. The following embodiments are,therefore, to be construed as merely illustrative, and not limitative ofthe remainder of the disclosure in any way whatsoever.

[0280] The disclosures of all patents, applications and publications,mentioned above and below, in particular U.S. Ser. No. 60/172,000, U.S.Ser. No. 60/176,083, U.S. Ser. No. 60/177,332, U.S. Ser. No. 60/178,572,U.S. Ser. No. 60/179,758, and U.S. Ser. No. 60/181,625, are expresslyincorporated by reference herein.

EXAMPLES

[0281] I. Construction of cDNA Libraries

[0282] Incyte cDNAs were derived from cDNA libraries described in theLIFESEQ GOLD database (Incyte Genomics, Palo Alto Calif.) and shown inTable 4, column 5. Some tissues were homogenized and lysed inguanidinium isothiocyanate, while others were homogenized and lysed inphenol or in a suitable mixture of denaturants, such as TRIZOL (LifeTechnologies), a monophasic solution of phenol and guanidineisothiocyanate. The resulting lysates were centrifuged over CsClcushions or extracted with chloroform. RNA was precipitated from thelysates with either isopropanol or sodium acetate and ethanol, or byother routine methods.

[0283] Phenol extraction and precipitation of RNA were repeated asnecessary to increase RNA purity. In some cases, RNA was treated withDNase. For most libraries, poly(A)+ RNA was isolated using oligod(T)-coupled paramagnetic particles (Promega), OLIGOTEX latex particles(QIAGEN, Chatsworth Calif.), or an OLIGOTEX mRNA purification kit(QIAGEN). Alternatively, RNA was isolated directly from tissue lysatesusing other RNA isolation kits, e.g., the POLY(A)PURE mRNA purificationkit (Ambion, Austin Tex.).

[0284] In some cases, Stratagene was provided with RNA and constructedthe corresponding cDNA libraries. Otherwise, cDNA was synthesized andcDNA libraries were constructed with the UNIZAP vector system(Stratagene) or SUPERSCRIPT plasmid system (Life Technologies), usingthe recommended procedures or similar methods known in the art. (See,e.g., Ausubel, 1997, supra, units 5.1-6.6.) Reverse transcription wasinitiated using oligo d(T) or random primers. Synthetic oligonucleotideadapters were ligated to double stranded cDNA, and the cDNA was digestedwith the appropriate restriction enzyme or enzymes. For most libraries,the cDNA was size-selected (300-1000 bp) using SEPHACRYL S1000,SEPHAROSE CL2B, or SEPHAROSE CL4B column chromatography (AmershamPharmacia Biotech) or preparative agarose gel electrophoresis. cDNAswere ligated into compatible restriction enzyme sites of the polylinkerof a suitable plasmid, e.g., PBLUESCRIPT plasmid (Stratagene), PSPORT1plasmid (Life Technologies), PCDNA2.1 plasmid (Invitrogen, CarlsbadCalif.), or pINCY (Incyte Genomics, Palo Alto Calif.). Recombinantplasmids were transformed into competent E. coli cells includingXL1-Blue, XL1-BlueMRF, or SOLR from Stratagene or DH5α, DH10B, orEletroMAX DH10B from Life Technologies.

[0285] II. Isolation of cDNA Clones

[0286] Plasmids obtained as described in Example I were recovered fromhost cells by in vivo excision using the UNIZAP vector system(Stratagene) or by cell lysis. Plasmids were purified using at least oneof the following: a Magic or WIZARD Minipreps DNA purification system(Promega); an AGTC Miniprep purification kit (Edge Biosystems,Gaithersburg Md.); and QIAWELL 8 Plasmid, QIAWELL 8 Plus Plasmid,QIAWELL 8 Ultra Plasmid purification systems or the R.E.A.L. PREP 96plasmid purification kit from QIAGEN. Following precipitation, plasmidswere resuspended in 0.1 ml of distilled water and stored, with orwithout lyophilization, at 4° C.

[0287] Alternatively, plasmid DNA was amplified from host cell lysatesusing direct link PCR in a high-throughput format (Rao, V.B. (1994)Anal. Biochem 216:1-14). Host cell lysis and thermal cycling steps werecarried out in a single reaction mixture. Samples were processed andstored in 384-well plates, and the concentration of amplified plasmidDNA was quantified fluorometrically using PICOGREEN dye (MolecularProbes, Eugene Oreg.) and a FLUOROSKAN II fluorescence scanner(Labsystems Oy, Helsinki, Finland).

[0288] III. Sequencing and Analysis

[0289] Incyte cDNA recovered in plasmids as described in Example II weresequenced as follows. Sequencing reactions were processed using standardmethods or high-throughput instrumentation such as the ABI CATALYST 800(PE Biosystems) thermal cycler or the PTC-200 thermal cycler (MJResearch) in conjunction with the HYDRA microdispenser (RobbinsScientific) or the MICROLAB 2200 (Hamilton) liquid transfer system. cDNAsequencing reactions were prepared using reagents provided by AmershamPharmacia Biotech or supplied in ABI sequencing kits such as the ABIPRISM BIGDYE Terminator cycle sequencing ready reaction kit (PEBiosystems). Electrophoretic separation of cDNA sequencing reactions anddetection of labeled polynucleotides were carried out using the MEGABACE1000 DNA sequencing system (Molecular Dynamics); the ABI PRISM 373 or377 sequencing system (PE Biosystems) in conjunction with standard ABIprotocols and base calling software; or other sequence analysis systemsknown in the art. Reading frames within the cDNA sequences wereidentified using standard methods (reviewed in Ausubel, 1997, supra,unit 7.7). Some of the cDNA sequences were selected for extension usingthe techniques disclosed in Example VIII.

[0290] The polynucleotide sequences derived from Incyte cDNAs werevalidated by removing vector, linker, and poly(A) sequences and bymasking ambiguous bases, using algorithms and programs based on BLAST,dynamic programming, and dinucleotide nearest neighbor analysis. TheIncyte cDNA sequences or translations thereof were then queried againsta selection of public databases such as the GenBank primate, rodent,mammalian, vertebrate, and eukaryote databases, and BLOCKS, PRINTS,DOMO, PRODOM, and hidden Markov model (HMM)-based protein familydatabases such as PFAM. (HMM is a probabilistic approach which analyzesconsensus primary structures of gene families. See, for example, Eddy,S. R. (1996) Curr. Opin. Struct. Biol. 6:361-365.) The queries wereperformed using programs based on BLAST, FASTA, BLIMPS, and HMMR. TheIncyte cDNA sequences were assembled to produce full lengthpolynucleotide sequences. Alternatively, GenBank cDNAs, GenBank ESTs,stitched sequences, stretched sequences, or Genscan-predicted codingsequences (see Examples IV and V) were used to extend Incyte cDNAassemblages to full length. Assembly was performed using programs basedon Phred, Phrap, and Consed, and cDNA assemblages were screened for openreading frames using programs based on GeneMark, BLAST, and FASTA. Thefull length polynucleotide sequences were translated to derive thecorresponding full length polypeptide sequences which were subsequentlyanalyzed by querying against databases such as the GenBank proteindatabases (genpept), SwissProt, BLOCKS, PRINTS, DOMO, PRODOM, Prosite,and hidden Markov model (HMM)-based protein family databases such asPFAM. Full length polynucleotide sequences are also analyzed usingMACDNASIS PRO software (Hitachi Software Engineering, South SanFrancisco Calif.) and LASERGENE software (DNASTAR). Polynucleotide andpolypeptide sequence alignments are generated using default parametersspecified by the CLUSTAL algorithm as incorporated into the MEGALIGNmultisequence alignment program (DNASTAR), which also calculates thepercent identity between aligned sequences.

[0291] Table 7 summarizes the tools, programs, and algorithms used forthe analysis and assembly of Incyte cDNA and full length sequences andprovides applicable descriptions, references, and threshold parameters.The first column of Table 7 shows the tools, programs, and algorithmsused, the second column provides brief descriptions thereof, the thirdcolumn presents appropriate references, all of which are incorporated byreference herein in their entirety, and the fourth column presents,where applicable, the scores, probability values, and other parametersused to evaluate the strength of a match between two sequences (thehigher the score or the lower the probability value, the greater theidentity between two sequences).

[0292] The programs described above for the assembly and analysis offull length polynucleotide and polypeptide sequences were also used toidentify polynucleotide sequence fragments from SEQ ID NO:28-54.Fragments from about 20 to about 4000 nucleotides which are useful inhybridization and amplification technologies are described in Table 4,column 4.

[0293] IV. Identification and Editing of Coding Sequences from GenomicDNA

[0294] Putative transporters and ion channels were initially identifiedby running the Genscan gene identification program against publicgenomic sequence databases (e.g., gbpri and gbhtg). Genscan is ageneral-purpose gene identification program which analyzes genomic DNAsequences from a variety of organisms (See Burge, C. and S. Karlin(1997) J. Mol. Biol. 268:78-94, and Burge, C. and S. Karlin (1998) Curr.Opin. Struct Biol. 8:346-354). The program concatenates predicted exonsto form an assembled cDNA sequence extending from a methionine to a stopcodon. The output of Genscan is a FASTA database of polynucleotide andpolypeptide sequences. The maximum range of sequence for Genscan toanalyze at once was set to 30 kb. To determine which of these Genscanpredicted cDNA sequences encode transporters and ion channels, theencoded polypeptides were analyzed by querying against PFAM models fortransporters and ion channels. Potential transporters and ion channelswere also identified by homology to Incyte cDNA sequences that had beenannotated as transporters and ion channels. These selectedGenscan-predicted sequences were then compared by BLAST analysis to thegenpept and gbpri public databases. Where necessary, theGenscan-predicted sequences were then edited by comparison to the topBLAST hit from genpept to correct errors in the sequence predicted byGenscan, such as extra or omitted exons. BLAST analysis was also used tofind any Incyte cDNA or public cDNA coverage of the Genscan-predictedsequences, thus providing evidence for transcription. When Incyte cDNAcoverage was available, this information was used to correct or confirmthe Genscan predicted sequence. Full length polynucleotide sequenceswere obtained by assembling Genscan-predicted coding sequences withIncyte cDNA sequences and/or public cDNA sequences using the assemblyprocess described in Example III. Alternatively, full lengthpolynucleotide sequences were derived entirely from edited or uneditedGenscan-predicted coding sequences.

[0295] V. Assembly of Genomic Sequence Data with cDNA Sequence Data

[0296] “Stitched” Sequences

[0297] Partial cDNA sequences were extended with exons predicted by theGenscan gene identification program described in Example IV. PartialcDNAs assembled as described in Example m were mapped to genomic DNA andparsed into clusters containing related cDNAs and Genscan exonpredictions from one or more genomic sequences. Each cluster wasanalyzed using an algorithm based on graph theory and dynamicprogramming to integrate cDNA and genomic information, generatingpossible splice variants that were subsequently confirmed, edited, orextended to create a full length sequence. Sequence intervals in whichthe entire length of the interval was present on more than one sequencein the cluster were identified, and intervals thus identified wereconsidered to be equivalent by transitivity. For example, if an intervalwas present on a cDNA and two genomic sequences, then all threeintervals were considered to be equivalent. This process allowsunrelated but consecutive genomic sequences to be brought together,bridged by cDNA sequence. Intervals thus identified were then “stitched”together by the stitching algorithm in the order that they appear alongtheir parent sequences to generate the longest possible sequence, aswell as sequence variants. Linkages between intervals which proceedalong one type of parent sequence (cDNA to cDNA or genomic sequence togenomic sequence) were given preference over linkages which changeparent type (cDNA to genomic sequence). The resultant stitched sequenceswere translated and compared by BLAST analysis to the genpept and gbpripublic databases. Incorrect exons predicted by Genscan were corrected bycomparison to the top BLAST hit from genpept. Sequences were furtherextended with additional cDNA sequences, or by inspection of genomicDNA, when necessary.

[0298] “Stretched” Sequences

[0299] Partial DNA sequences were extended to full length with analgorithm based on BLAST analysis. First, partial cDNAs assembled asdescribed in Example III were queried against public databases such asthe GenBank primate, rodent, mammalian, vertebrate, and eukaryotedatabases using the BLAST program. The nearest GenBank protein homologwas then compared by BLAST analysis to either Incyte cDNA sequences orGenScan exon predicted sequences described in Example IV. A chimericprotein was generated by using the resultant high-scoring segment pairs(HSPs) to map the translated sequences onto the GenBank protein homolog.Insertions or deletions may occur in the chimeric protein with respectto the original GenBank protein homolog. The GenBank protein homolog,the chimeric protein, or both were used as probes to search forhomologous genomic sequences from the public human genome databases.Partial DNA sequences were therefore “stretched” or extended by theaddition of homologous genomic sequences. The resultant stretchedsequences were examined to determine whether it contained a completegene.

[0300] VI. Chromosomal Mapping of TRICH Encoding Polynucleotides

[0301] The sequences which were used to assemble SEQ ID NO:28-54 werecompared with sequences from the Incyte LIFESEQ database and publicdomain databases using BLAST and other implementations of theSmith-Waterman algorithm. Sequences from these databases that matchedSEQ ID NO:28-54 were assembled into clusters of contiguous andoverlapping sequences using assembly algorithms such as Phrap (Table 7).Radiation hybrid and genetic mapping data available from publicresources such as the Stanford Human Genome Center (SHGC), WhiteheadInstitute for Genome Research (WIGR), and Généthon were used todetermine if any of the clustered sequences had been previously mapped.Inclusion of a mapped sequence in a cluster resulted in the assignmentof all sequences of that cluster, including its particular SEQ ID NO:,to that map location.

[0302] Map locations are represented by ranges, or intervals, or humanchromosomes. The map position of an interval, in centiMorgans, ismeasured relative to the terminus of the chromosome's p-arm. (ThecentiMorgan (cM) is a unit of measurement based on recombinationfrequencies between chromosomal markers. On average, 1 cM is roughlyequivalent to 1 megabase (Mb) of DNA in humans, although this can varywidely due to hot and cold spots of recombination.) The cM distances arebased on genetic markers mapped by Généthon which provide boundaries forradiation hybrid markers whose sequences were included in each of theclusters. Human genome maps and other resources available to the public,such as the NCBI “GeneMap'99” World Wide Web site(http://www.ncbi.nlm.nihgov/genemap/), can be employed to determine ifpreviously identified disease genes map within or in proximity to theintervals indicated above.

[0303] VII. Analysis of Polynucleotide Expression

[0304] Northern analysis is a laboratory technique used to detect thepresence of a transcript of a gene and involves the hybridization of alabeled nucleotide sequence to a membrane on which RNAs from aparticular cell type or tissue have been bound. (See, e.g., Sambrook,supra, ch 7; Ausubel (1995) supra, ch. 4 and 16.)

[0305] Analogous computer techniques applying BLAST were used to searchfor identical or related molecules in cDNA databases such as GenBank orLIFESEQ (Incyte Genomics). This analysis is much faster than multiplemembrane-based hybridizations. In addition, the sensitivity of thecomputer search can be modified to determine whether any particularmatch is categorized as exact or similar. The basis of the search is theproduct score, which is defined as:$\frac{\text{BLAST~~Score} \times \text{Percent~~Identity}}{5 \times \quad \text{minimum}\quad \left\{ {{{length}\left( {{Seq}.\quad 1} \right)},{{length}\left( {{Seq}.\quad 2} \right)}} \right\}}$

[0306] The product score takes into account both the degree ofsimilarity between two sequences and the length of the sequence match.The product score is a normalized value between 0 and 100, and iscalculated as follows: the BLAST score is multiplied by the percentnucleotide identity and the product is divided by (5 times the length ofthe shorter of the two sequences). The BLAST score is calculated byassigning a score of +5 for every base that matches in a high-scoringsegment pair (HSP), and −4 for every mismatch. Two sequences may sharemore than one HSP (separated by gaps). If there is more than one HSP,then the pair with the highest BLAST score is used to calculate theproduct score. The product score represents a balance between fractionaloverlap and quality in a BLAST alignment. For example, a product scoreof 100 is produced only for 100% identity over the entire length of theshorter of the two sequences being compared. A product score of 70 isproduced either by 100% identity and 70% overlap at one end, or by 88%identity and 100% overlap at the other. A product score of 50 isproduced either by 100% identity and 50% overlap at one end, or 79%identity and 100% overlap.

[0307] Alternatively, polynucleotide sequences encoding TRICH areanalyzed with respect to the tissue sources from which they werederived. For example, some full length sequences are assembled, at leastin part, with overlapping Incyte cDNA sequences (see Example II). EachcDNA sequence is derived from a cDNA library constructed from a humantissue. Each human tissue is classified into one of the followingorgan/tissue categories: cardiovascular system; connective tissue;digestive system; embryonic structures; endocrine system; exocrineglands; genitalia, female; genitalia, male; germ cells; hemic and immunesystem; liver; musculoskeletal system; nervous system; pancreas;respiratory system; sense organs; skin; stomatognathic system;unclassified/mixed; or urinary tract. The number of libraries in eachcategory is counted and divided by the total number of libraries acrossall categories. Similarly, each human tissue is classified into one ofthe following disease/condition categories: cancer, cell line,developmental, inflammation, neurological, trauma, cardiovascular,pooled, and other, and the number of libraries in each category iscounted and divided by the total number of libraries across allcategories. The resulting percentages reflect the tissue- anddisease-specific expression of cDNA encoding TRICH. cDNA sequences andcDNA library/tissue information are found in the LIFESEQ GOLD database(Incyte Genomics, Palo Alto Calif.).

[0308] VIII. Extension of TRICH Encoding Polynucleotides

[0309] Full length polynucleotide sequences were also produced byextension of an appropriate fragment of the full length molecule usingoligonucleotide primers designed from this fragment. One primer wassynthesized to initiate 5′ extension of the known fragment, and theother primer was synthesized to initiate 3′ extension of the knownfragment. The initial primers were designed using OLIGO 4.06 software(National Biosciences), or another appropriate program, to be about 22to 30 nucleotides in length, to have a GC content of about 50% or more,and to anneal to the target sequence at temperatures of about 68° C. toabout 72° C. Any stretch of nucleotides which would result in hairpinstructures and primer-primer dimerizations was avoided.

[0310] Selected human cDNA libraries were used to extend the sequence.If more than one extension was necessary or desired, additional ornested sets of primers were designed.

[0311] High fidelity amplification was obtained by PCR using methodswell known in the art. PCR was performed in 96-well plates using thePTC-200 thermal cycler (MJ Research, Inc.). The reaction mix containedDNA template, 200 mmol of each primer, reaction buffer containing Mg⁺,(NH₄)₂SO₄, and 2-mercaptoethanol, Taq DNA polymerase (Amersham PharmaciaBiotech), ELONGASE enzyme (Life Technologies), and Pfu DNA polymerase(Stratagene), with the following parameters for primer pair PCI A andPCI B: Step 1: 94° C., 3 min; Step 2: 94° C., 15 sec; Step 3: 60° C., 1min; Step 4: 68° C., 2 min; Step 5: Steps 2, 3, and 4 repeated 20 times;Step 6: 68° C., 5 min; Step 7: storage at 4° C. In the alternative, theparameters for primer pair T7 and SK+ were as follows: Step 1: 94° C., 3min; Step 2: 94° C., 15 sec; Step 3: 57° C., 1 min; Step 4: 68° C., 2min; Step 5: Steps 2, 3, and 4 repeated 20 times; Step 6: 68° C., 5 min;Step 7: storage at 4° C.

[0312] The concentration of DNA in each well was determined bydispensing 100 μl PICOGREEN quantitation reagent (0.25% (v/v) PICOGREEN;Molecular Probes, Eugene Oreg.) dissolved in 1×TE and 0.5 μl ofundiluted PCR product into each well of an opaque fluorimeter plate(Corning Costar, Acton Mass.), allowing the DNA to bind to the reagent.The plate was scanned in a Fluoroskan II (Labsystems Oy, Helsinki,Finland) to measure the fluorescence of the sample and to quantify theconcentration of DNA. A 5 μl to 10 μl aliquot of the reaction mixturewas analyzed by electrophoresis on a 1% agarose gel to determine whichreactions were successful in extending the sequence.

[0313] The extended nucleotides were desalted and concentrated,transferred to 384-well plates, digested with CviJI cholera virusendonuclease (Molecular Biology Research, Madison Wis.), and sonicatedor sheared prior to religation into pUC 18 vector (Amersham PharmaciaBiotech). For shotgun sequencing, the digested nucleotides wereseparated on low concentration (0.6 to 0.8%) agarose gels, fragmentswere excised, and agar digested with Agar ACE (Promega). Extended cloneswere religated using T4 ligase (New England Biolabs, Beverly Mass.) intopUC 18 vector (Amersham Pharmacia Biotech), treated with Pfu DNApolymerase (Stratagene) to fill-in restriction site overhangs, andtransfected into competent E. coli cells. Transformed cells wereselected on antibiotic-containing media, and individual colonies werepicked and cultured overnight at 37° C. in 384-well plates in LB/2×carbliquid media.

[0314] The cells were lysed, and DNA was amplified by PCR using Taq DNApolymerase (Amersham Pharmacia Biotech) and Pfu DNA polymerase(Stratagene) with the following parameters: Step 1: 94° C., 3 min; Step2: 94° C., 15 sec; Step 3: 60° C., 1 min; Step 4: 72° C., 2 min; Step 5:steps 2, 3, and 4 repeated 29 times; Step 6: 72° C., S min; Step 7:storage at 4° C. DNA was quantified by PICOGREEN reagent (MolecularProbes) as described above. Samples with low DNA recoveries werereamplified using the same conditions as described above. Samples werediluted with 20% dimethysulfoxide (1:2, v/v), and sequenced usingDYENAMIC energy transfer sequencing primers and the DYENAMIC DIRECT kit(Amersham Pharmacia Biotech) or the ABI PRISM BIGDYE Terminator cyclesequencing ready reaction kit (PE Biosystems).

[0315] In like manner, full length polynucleotide sequences are verifiedusing the above procedure or are used to obtain 5′ regulatory sequencesusing the above procedure along with oligonucleotides designed for suchextension, and an appropriate genomic library.

[0316] IX. Labeling and Use of Individual Hybridization Probes

[0317] Hybridization probes derived from SEQ ID NO:28-54 are employed toscreen cDNAs, genomic DNAs, or mRNAs. Although the labeling ofoligonucleotides, consisting of about 20 base pairs, is specificallydescribed, essentially the same procedure is used with larger nucleotidefragments. Oligonucleotides are designed using state-of-the-art softwaresuch as OLIGO 4.06 software (National Biosciences) and labeled bycombining 50 pmol of each oligomer, 250 μCi of [γ-³²P] adenosinetriphosphate (Amersham Pharmacia Biotech), and T4 polynucleotide kinase(DuPont NEN, Boston Mass.). The labeled oligonucleotides aresubstantially purified using a SEPHADEX G-25 superfine size exclusiondextran bead column (Amersham Pharmacia Biotech). An aliquot containing10⁷ counts per minute of the labeled probe is used in a typicalmembrane-based hybridization analysis of human genomic DNA digested withone of the following endonucleases: Ase I, Bgl II, Eco RI, Pst I, Xba I,or Pvu II (DuPont NEN).

[0318] The DNA from each digest is fractionated on a 0.7% agarose geland transferred to nylon membranes (Nytran Plus, Schleicher & Schuell,Durham N.H.). Hybridization is carried out for 16 hours at 40° C. Toremove nonspecific signals, blots are sequentially washed at roomtemperature under conditions of up to, for example, 0.1×saline sodiumcitrate and 0.5% sodium dodecyl sulfate. Hybridization patterns arevisualized using autoradiography or an alternative imaging means andcompared.

[0319] X. Microarrays

[0320] The linkage or synthesis of array elements upon a microarray canbe achieved utilizing photolithography, piezoelectric printing (ink-jetprinting, See, e.g., Baldeschweiler, supra.), mechanical microspottingtechnologies, and derivatives thereof. The substrate in each of theaforementioned technologies should be uniform and solid with anon-porous surface (Schena (1999), supra). Suggested substrates includesilicon, silica, glass slides, glass chips, and silicon wafers.Alternatively, a procedure analogous to a dot or slot blot may also beused to arrange and link elements to the surface of a substrate usingthermal, UV, chemical, or mechanical bonding procedures. A typical arraymay be produced using available methods and machines well known to thoseof ordinary skill in the art and may contain any appropriate number ofelements. (See, e.g., Schena, M. et al. (1995) Science 270:467-470;Shalon, D. et al. (1996) Genome Res. 6:639-645; Marshall, A. and J.Hodgson (1998) Nat. Biotechnol. 16:27-31.)

[0321] Full length cDNAs, Expressed Sequence Tags (ESTs), or fragmentsor oligomers thereof may comprise the elements of the microarray.Fragments or oligomers suitable for hybridization can be selected usingsoftware well known in the art such as LASERGENE software (DNASTAR). Thearray elements are hybridized with polynucleotides in a biologicalsample. The polynucleotides in the biological sample are conjugated to afluorescent label or other molecular tag for ease of detection. Afterhybridization, nonhybridized nucleotides from the biological sample areremoved, and a fluorescence scanner is used to detect hybridization ateach array element. Alternatively, laser desorbtion and massspectrometry may be used for detection of hybridization. The degree ofcomplementarity and the relative abundance of each polynucleotide whichhybridizes to an element on the microarray may be assessed. In oneembodiment, microarray preparation and usage is described in detailbelow.

[0322] Tissue or Cell Sample Preparation

[0323] Total RNA is isolated from tissue samples using the guanidiniumthiocyanate method and poly(A)⁺ RNA is purified using the oligo-(dT)cellulose method. Each poly(A)⁺ RNA sample is reverse transcribed usingMMLV reverse-transcriptase, 0.05 pg/μl oligo-(dT) primer (21mer),1×first strand buffer, 0.03 units/μl RNase inhibitor, 500 μM dATP, 500μM dGTP, 500 μM dTTP, 40 μM dCTP, 40 μM dCTP-Cy3 (BDS) or dCTP-Cy5(Amersham Pharmacia Biotech). The reverse transcription reaction isperformed in a 25 ml volume containing 200 ng poly(A)⁺ RNA withGEMBRIGHT kits (Incyte). Specific control poly(A)⁺ RNAs are synthesizedby in vitro transcription from non-coding yeast genomic DNA. Afterincubation at 37° C. for 2 hr, each reaction sample (one with Cy3 andanother with Cy5 labeling) is treated with 2.5 ml of 0.5M sodiumhydroxide and incubated for 20 minutes at 85° C. to the stop thereaction and degrade the RNA. Samples are purified using two successiveCHROMA SPIN 30 gel filtration spin columns (CLONTECH Laboratories, Inc.(CLONTECH), Palo Alto Calif.) and after combining, both reaction samplesare ethanol precipitated using 1 ml of glycogen (1 mg/ml), 60 ml sodiumacetate, and 300 ml of 100% ethanol. The sample is then dried tocompletion using a SpeedVAC (Savant Instruments Inc., Holbrook N.Y.) andresuspended in 14 μl 5×SSC/0.2% SDS.

[0324] Microarray Preparation

[0325] Sequences of the present invention are used to generate arrayelements. Each array element is amplified from bacterial cellscontaining vectors with cloned cDNA inserts. PCR amplification usesprimers complementary to the vector sequences flanking the cDNA insert.Array elements are amplified in thirty cycles of PCR from an initialquantity of 1-2 ng to a final quantity greater than 5 μg. Amplifiedarray elements are then purified using SEPHACRYL-400 (Amersham PharmaciaBiotech).

[0326] Purified array elements are immobilized on polymer-coated glassslides. Glass microscope slides (Corning) are cleaned by ultrasound in0.1% SDS and acetone, with extensive distilled water washes between andafter treatments. Glass slides are etched in 4% hydrofluoric acid (VWRScientific Products Corporation (VWR), West Chester Pa.), washedextensively in distilled water, and coated with 0.05% aminopropyl silane(Sigma) in 95% ethanol. Coated slides are cured in a 110° C. oven.

[0327] Array elements are applied to the coated glass substrate using aprocedure described in U.S. Pat. No. 5,807,522, incorporated herein byreference. 1 μl of the array element DNA, at an average concentration of100 ng/μl, is loaded into the open capillary printing element by ahigh-speed robotic apparatus. The apparatus then deposits about 5 nl ofarray element sample per slide.

[0328] Microarrays are UV-crosslinked using a STRATALINKERUV-crosslinker (Stratagene). Microarrays are washed at room temperatureonce in 0.2% SDS and three times in distilled water. Non-specificbinding sites are blocked by incubation of microarrays in 0.2% casein inphosphate buffered saline (PBS) (Tropix, Inc., Bedford Mass.) for 30minutes at 60° C. followed by washes in 0.2% SDS and distilled water asbefore.

[0329] Hybridization

[0330] Hybridization reactions contain 9 μl of sample mixture consistingof 0.2 μg each of Cy3 and Cy5 labeled cDNA synthesis products in 5×SSC,0.2% SDS hybridization buffer. The sample mixture is heated to 65° C.for 5 minutes and is aliquoted onto the microarray surface and coveredwith an 1.8 cm² coverslip. The arrays are transferred to a waterproofchamber having a cavity just slightly larger than a microscope slide.The chamber is kept at 100% humidity internally by the addition of 140μ, of 5×SSC in a corner of the chamber. The chamber containing thearrays is incubated for about 6.5 hours at 60° C. The arrays are washedfor 10 min at 45° C. in a first wash buffer (1×SSC, 0.1% SDS), threetimes for 10 minutes each at 45° C. in a second wash buffer (0.1×SSC),and dried.

[0331] Detection

[0332] Reporter-labeled hybridization complexes are detected with amicroscope equipped with an Innova 70 mixed gas 10 W laser (Coherent,Inc., Santa Clara Calif.) capable of generating spectral lines at 488 nmfor excitation of Cy3 and at 632 nm for excitation of Cy5. Theexcitation laser light is focused on the array using a 20× microscopeobjective (Nikon, Inc., Melville N.Y.). The slide containing the arrayis placed on a computer-controlled X-Y stage on the microscope andraster-scanned past the objective. The 1.8 cm×1.8 cm array used in thepresent example is scanned with a resolution of 20 micrometers.

[0333] In two separate scans, a mixed gas multiline laser excites thetwo fluorophores sequentially. Emitted light is split, based onwavelength, into two photomultiplier tube detectors (PMT R1477,Hamamatsu Photonics Systems, Bridgewater N.J.) corresponding to the twofluorophores. Appropriate filters positioned between the array and thephotomultiplier tubes are used to filter the signals. The emissionmaxima of the fluorophores used are 565 nm for Cy3 and 650 nm for Cy5.Each array is typically scanned twice, one scan per fluorophore usingthe appropriate filters at the laser source, although the apparatus iscapable of recording the spectra from both fluorophores simultaneously.

[0334] The sensitivity of the scans is typically calibrated using thesignal intensity generated by a cDNA control species added to the samplemixture at a known concentration. A specific location on the arraycontains a complementary DNA sequence, allowing the intensity of thesignal at that location to be correlated with a weight ratio ofhybridizing species of 1:100,000. When two samples from differentsources (e.g., representing test and control cells), each labeled with adifferent fluorophore, are hybridized to a single array for the purposeof identifying genes that are differentially expressed, the calibrationis done by labeling samples of the calibrating cDNA with the twofluorophores and adding identical amounts of each to the hybridizationmixture.

[0335] The output of the photomultiplier tube is digitized using a12-bit RTI-835H analog-to-digital (A/D) conversion board (AnalogDevices, Inc., Norwood Mass.) installed in an IBM-compatible PCcomputer. The digitized data are displayed as an image where the signalintensity is mapped using a linear 20-color transformation to apseudocolor scale ranging from blue (low signal) to red (high signal).The data is also analyzed quantitatively. Where two differentfluorophores are excited and measured simultaneously, the data are firstcorrected for optical crosstalk (due to overlapping emission spectra)between the fluorophores using each fluorophore's emission spectrum.

[0336] A grid is superimposed over the fluorescence signal image suchthat the signal from each spot is centered in each element of the grid.The fluorescence signal within each element is then integrated to obtaina numerical value corresponding to the average intensity of the signal.The software used for signal analysis is the GEMTOOLS gene expressionanalysis program (Incyte). XI. Complementary Polynucleotides

[0337] Sequences complementary to the TRICH-encoding sequences, or anyparts thereof, are used to detect, decrease, or inhibit expression ofnaturally occurring TRICH. Although use of oligonucleotides comprisingfrom about 15 to 30 base pairs is described, essentially the sameprocedure is used with smaller or with larger sequence fragments.Appropriate oligonucleotides are designed using OLIGO 4.06 software(National Biosciences) and the coding sequence of TRICH. To inhibittranscription, a complementary oligonucleotide is designed from the mostunique 5′ sequence and used to prevent promoter binding to the codingsequence. To inhibit translation, a complementary oligonucleotide isdesigned to prevent ribosomal binding to the TRICH-encoding transcript.

[0338] XII. Expression of TRICH

[0339] Expression and purification of TRICH is achieved using bacterialor virus-based expression systems. For expression of TRICH in bacteria,cDNA is subcloned into an appropriate vector containing an antibioticresistance gene and an inducible promoter that directs high levels ofcDNA transcription. Examples of such promoters include, but are notlimited to, the trp-lac (tac) hybrid promoter and the T5 or T7bacteriophage promoter in conjunction with the lac operator regulatoryelement. Recombinant vectors are transformed into suitable bacterialhosts, e.g., BL21(DE3). Antibiotic resistant bacteria express TRICH uponinduction with isopropyl beta-D-thiogalactopyranoside (IPTG). Expressionof TRICH in eukaryotic cells is achieved by infecting insect ormamnalian cell lines with recombinant Autographica californica nuclearpolyhedrosis virus (AcMNPV), commonly known as baculovirus. Thenonessential polyhedrin gene of baculovirus is replaced with cDNAencoding TRICH by either homologous recombination or bacterial-mediatedtransposition involving transfer plasmid intermediates. Viralinfectivity is maintained and the strong polyhedrin promoter drives highlevels of cDNA transcription. Recombinant baculovirus is used to infectSpodoptera frugiperda (Sf9) insect cells in most cases, or humanhepatocytes, in some cases. Infection of the latter requires additionalgenetic modifications to baculovirus. (See Engelhard, E. K. et al.(1994) Proc. Natl. Acad. Sci. USA 91:3224-3227; Sandig, V. et al. (1996)Hum. Gene Ther. 7:1937-1945.)

[0340] In most expression systems, TRICH is synthesized as a fusionprotein with, e.g., glutathione S-transferase (GST) or a peptide epitopetag, such as FLAG or 6-His, permitting rapid, single-step,affinity-based purification of recombinant fusion protein from crudecell lysates. GST, a 26-kilodalton enzyme from Schistosoma japonicum,enables the purification of fusion proteins on immobilized glutathioneunder conditions that maintain protein activity and antigenicity(Amersham Pharmacia Biotech). Following purification, the GST moiety canbe proteolytically cleaved from TRICH at specifically engineered sites.FLAG, an 8-amino acid peptide, enables immunoaffinity purification usingcommercially available monoclonal and polyclonal anti-FLAG antibodies(Eastman Kodak). 6-His, a stretch of six consecutive histidine residues,enables purification on metal-chelate resins (QIAGEN). Methods forprotein expression and purification are discussed in Ausubel (1995,supra, ch. 10 and 16). Purified TRICH obtained by these methods can beused directly in the assays shown in Examples XVI, XVII, and XVIII,where applicable.

[0341] XIII. Functional Assays

[0342] TRICH function is assessed by expressing the sequences encodingTRICH at physiologically elevated levels in mammalian cell culturesystems. cDNA is subcloned into a mammalian expression vector containinga strong promoter that drives high levels of cDNA expression. Vectors ofchoice include PCMV SPORT (Life Technologies) and PCR3.1 (Invitrogen,Carlsbad Calif.), both of which contain the cytomegalovirus promoter.5-10 μg of recombinant vector are transiently transfected into a humancell line, for example, an endothelial or hematopoietic cell line, usingeither liposome formulations or electroporation. 1-2 μg of an additionalplasmid containing sequences encoding a marker protein areco-transfected. Expression of a marker protein provides a means todistinguish transfected cells from nontransfected cells and is areliable predictor of cDNA expression from the recombinant vector.Marker proteins of choice include, e.g., Green Fluorescent Protein (GFP;Clontech), CD64, or a CD64-GFP fusion protein. Flow cytometry (FCM), anautomated, laser optics-based technique, is used to identify transfectedcells expressing GFP or CD64-GFP and to evaluate the apoptotic state ofthe cells and other cellular properties. FCM detects and quantifies theuptake of fluorescent molecules that diagnose events preceding orcoincident with cell death. These events include changes in nuclear DNAcontent as measured by staining of DNA with propidium iodide; changes incell size and granularity as measured by forward light scatter and 90degree side light scatter; down-regulation of DNA synthesis as measuredby decrease in bromodeoxyuridine uptake; alterations in expression ofcell surface and intracellular proteins as measured by reactivity withspecific antibodies; and alterations in plasma membrane composition asmeasured by the binding of fluorescein-conjugated Annexin V protein tothe cell surface. Methods in flow cytometry are discussed in Ormerod, M.G. (1994) Flow Cytometry, Oxford, New York N.Y.

[0343] The influence of TRICH on gene expression can be assessed usinghighly purified populations of cells transfected with sequences encodingTRICH and either CD64 or CD64-GFP. CD64 and CD64-GFP are expressed onthe surface of transfected cells and bind to conserved regions of humanimmunoglobulin G (IgG). Transfected cells are efficiently separated fromnontransfected cells using magnetic beads coated with either human IgGor antibody against CD64 (DYNAL, Lake Success N.Y.). mRNA can bepurified from the cells using methods well known by those of skill inthe art. Expression of mRNA encoding TRICH and other genes of interestcan be analyzed by northern analysis or microarray techniques.

[0344] XIV. Production of TRICH Specific Antibodies

[0345] TRICH substantially purified using polyacrylamide gelelectrophoresis (PAGE; see, e.g., Harrington, M. G. (1990) MethodsEnzymol. 182:488-495), or other purification techniques, is used toimmunize rabbits and to produce antibodies using standard protocols.

[0346] Alternatively, the TRICH amino acid sequence is analyzed usingLASERGENE software (DNASTAR) to determine regions of highimmunogenicity, and a corresponding oligopeptide is synthesized and usedto raise antibodies by means known to those of skill in the art. Methodsfor selection of appropriate epitopes, such as those near the C-terminusor in hydrophilic regions are well described in the art. (See, e.g.,Ausubel, 1995, supra, ch. 11.)

[0347] Typically, oligopeptides of about 15 residues in length aresynthesized using an ABI 431A peptide synthesizer (PE Biosystems) usingFMOC chemistry and coupled to KLH (Sigma-Aldrich, St Louis Mo.) byreaction with N-maleinidobenzoyl-N-hydroxysuccinimide ester (MBS) toincrease immunogenicity. (See, e.g., Ausubel, 1995, supra.) Rabbits areimmunized with the oligopeptide-KLH complex in complete Freund'sadjuvant. Resulting antisera are tested for antipeptide and anti-TRICHactivity by, for example, binding the peptide or TRICH to a substrate,blocking with 1% BSA, reacting with rabbit antisera, washing, andreacting with radio-iodinated goat anti-rabbit IgG.

[0348] XV. Purification of Naturally Occurring TRICH Using SpecificAntibodies

[0349] Naturally occurring or recombinant TRICH is substantiallypurified by immunoaffinity chromatography using antibodies specific forTRICH. An immunoaffinity column is constructed by covalently couplinganti-TRICH antibody to an activated chromatographic resin, such asCNBr-activated SEPHAROSE (Amersham Pharmacia Biotech). After thecoupling, the resin is blocked and washed according to themanufacturer's instructions.

[0350] Media containing TRICH are passed over the immunoaffinity column,and the column is washed under conditions that allow the preferentialabsorbance of TRICH (e.g., high ionic strength buffers in the presenceof detergent). The column is eluted under conditions that disruptantibody/TRICH binding (e.g., a buffer of pH 2 to pH 3, or a highconcentration of a chaotrope, such as urea or thiocyanate ion), andTRICH is collected.

[0351] XVI. Identification of Molecules Which Interact with TRICH

[0352] Molecules which interact with TRICH may include transportersubstrates, agonists or antagonists, modulatory proteins such as Gβγproteins (Reimann, supra) or proteins involved in TRICH localization orclustering such as MAGUKs (Craven, supra). TRICH, or biologically activefragments thereof, are labeled with ¹²⁵I Bolton-Hunter reagent. (See,e.g., Bolton A. E. and W. M. Hunter (1973) Biochem. J. 133:529-539.)Candidate molecules previously arrayed in the wells of a multi-wellplate are incubated with the labeled TRICH, washed, and any wells withlabeled TRICH complex are assayed. Data obtained using differentconcentrations of TRICH are used to calculate values for the number,affinity, and association of TRICH with the candidate molecules.

[0353] Alternatively, proteins that interact with TRICH are isolatedusing the yeast 2-hybrid system (Fields, S. and O. Song (1989) Nature340:245-246). TRICH, or fragments thereof, are expressed as fusionproteins with the DNA binding domain of Ga14 or lexA and potentialinteracting proteins are expressed as fusion proteins with an activationdomain. Interactions between the TRICH fusion protein and thereconstitutes a transactivation function that is observed by expressionof a reporter gene. Yeast 2-hybrid systems are commercially available,and methods for use of the yeast 2-hybrid system with ion channelproteins are discussed in Niethammer, M. and M. Sheng (1998, Meth.Enzymol. 293:104-122).

[0354] TRICH may also be used in the PATHCALLING process (CuraGen Corp.,New Haven Conn.) which employs the yeast two-hybrid system in ahigh-throughput manner to determine all interactions between theproteins encoded by two large libraries of genes (Nandabalan, K. et al.(2000) U.S. Pat. No. 6,057,101).

[0355] Potential TRICH agonists or antagonists may be tested foractivation or inhibition of TRICH ion channel activity using the assaysdescribed in section XVIII.

[0356] XVII. Demonstration of TRICH Activity

[0357] Ion channel activity of TRICH is demonstrated using anelectrophysiological assay for ion conductance. TRICH can be expressedby transforming a mammalian cell line such as COS7, HeLa or CHO with aeukaryotic expression vector encoding TRICH. Eukaryotic expressionvectors are commercially available, and the techniques to introduce theminto cells are well known to those skilled in the art A second plasmidwhich expresses any one of a number of marker genes, such asβ-galactosidase, is co-transformed into the cells to allow rapididentification of those cells which have taken up and expressed theforeign DNA. The cells are incubated for 48-72 hours aftertransformation under conditions appropriate for the cell line to allowexpression and accumulation of TRICH and β-galactosidase.

[0358] Transformed cells expressing β-galactosidase are stained bluewhen a suitable colorimetric substrate is added to the culture mediaunder conditions that are well known in the art. Stained cells aretested for differences in membrane conductance by electrophysiologicaltechniques that are well known in the art. Untransformed cells, and/orcells transformed with either vector sequences alone or β-galactosidasesequences alone, are used as controls and tested in parallel. Cellsexpressing TRICH will have higher anion or cation conductance relativeto control cells. The contribution of TRICH to conductance can beconfirmed by incubating the cells using antibodies specific for TRICH.The antibodies will bind to the extracellular side of TRICH, therebyblocking the pore in the ion channel, and the associated conductance.

[0359] Alternatively, ion channel activity of TRICH is measured ascurrent flow across a TRICH-containing Xenopus laevis oocyte membraneusing the two-electrode voltage-clamp technique (Ishi et al., supra;Jegla, T. and L. Salkoff (1997) J. Neurosci. 17:3244). TRICH issubcloned into an appropriate Xenopus oocyte expression vector, such aspBF, and 0.5-5 ng of mRNA is injected into mature stage IV oocytes.Injected oocytes are incubated at 18° C. for 1-5 days. Inside-outmacropatches are excised into an intracellular solution containing 116mM K-gluconate, 4 mM KCl, and 10 mM Hepes (pH 7.2). The intracellularsolution is supplemented with varying concentrations of the TRICHmediator, such as cAMP, cGMP, or Ca⁺² (in the form of CaCl₂), whereappropriate. Electrode resistance is set at 2-5 MΩ and electrodes arefilled with the intracellular solution lacking mediator. Experiments areperformed at room temperature from a holding potential of 0 mV. Voltageramps (2.5 s) from −100 to 100 mV are acquired at a sampling frequencyof 500 Hz. Current measured is proportional to the activity of TRICH inthe assay.

[0360] Transport activity of TRICH is assayed by measuring uptake oflabeled substrates into Xenopus laevis oocytes. Oocytes at stages V andVI are injected with TRICH mRNA (10 ng per oocyte) and incubated for 3days at 18° C. in OR2 medium (82.5 mM NaCl, 2.5 mM KCl, 1 mM CaCl₂, 1 mMMgCl₂, 1 mM Na₂HPO₄, 5 mM Hepes, 3.8 mM NaOH, 50 μg/ml gentamycin, pH7.8) to allow expression of TRICH. Oocytes are then transferred tostandard uptake medium (100 mM NaCl, 2 mM KCl, 1 mM CaCl₂, 10 mM MgCl₂,10 mM Hepes/Tris pH 7.5). Uptake of various substrates (e.g., aminoacids, sugars, drugs, ions, and neurotransmitters) is initiated byadding labeled substrate (e.g. radiolabeled with ³H, fluorescentlylabeled with rhodamine, etc.) to the oocytes. After incubating for 30minutes, uptake is terminated by washing the oocytes three times inNa⁺-free medium, measuring the incorporated label, and comparing withcontrols. TRICH activity is proportional to the level of internalizedlabeled substrate.

[0361] ATPase activity associated with TRICH can be measured byhydrolysis of radiolabeled ATP-[γ-³²P], separation of the hydrolysisproducts by chromatographic methods, and quantitation of the recovered³²P using a scintillation counter. The reaction mixture containsATP-[γ-³²P] and varying amounts of TRICH in a suitable buffer incubatedat 37° C. for a suitable period of time. The reaction is terminated byacid precipitation with trichloroacetic acid and then neutralized withbase, and an aliquot of the reaction mixture is subjected to membrane orfilter paper-based chromatography to separate the reaction products. Theamount of ³²P liberated is counted in a scintillation counter. Theamount of radioactivity recovered is proportional to the ATPase activityof TRICH in the assay.

[0362] XVIII. Identification of TRICH Agonists and Antagonists

[0363] TRICH is expressed in a eukaryotic cell line such as CHO (ChineseHamster Ovary) or HEK (Human Embryonic Kidney) 293. Ion channel activityof the transformed cells is measured in the presence and absence ofcandidate agonists or antagonists. Ion channel activity is assayed usingpatch clamp methods well known in the art or as described in ExampleXVII. Alternatively, ion channel activity is assayed using fluorescenttechniques that measure ion flux across the cell membrane (Velicelebi,G. et al. (1999) Meth Enzymol. 294:20-47; West, M. R. and C. R. Molloy(1996) Anal. Biochem. 241:51-58). These assays may be adapted forhigh-throughput screening using microplates. Changes in internal ionconcentration are measured using fluorescent dyes such as the Ca²⁺indicator Fluo-4 AM, sodium-sensitive dyes such as SBFI and sodiumgreen, or the Cl⁻indicator MQAE (all available from Molecular Probes) incombination with the FLIPR fluorimetric plate reading system (MolecularDevices). In a more generic version of this assay, changes in membranepotential caused by ionic flux across the plasma membrane are measuredusing oxonyl dyes such as DiBAC₄ (Molecular Probes). DiBAC₄ equilibratesbetween the extracellular solution and cellular sites according to thecellular membrane potential. The dye's fluorescence intensity is 20-foldgreater when bound to hydrophobic intracellular sites, allowingdetection of DiBAC₄ entry into the cell (Gonzalez, J. E. and P. A.Negulescu (1998) Curr. Opin. Biotechnol. 9:624-631). Candidate agonistsor antagonists may be selected from known ion channel agonists orantagonists, peptide libraries, or combinatorial chemical libraries.

[0364] Various modifications and variations of the described methods andsystems of the invention will be apparent to those skilled in the artwithout departing from the scope and spirit of the invention. Althoughthe invention has been described in connection with certain embodiments,it should be understood that the invention as claimed should not beunduly limited to such specific embodiments. Indeed, variousmodifications of the described modes for carrying out the inventionwhich are obvious to those skilled in molecular biology or relatedfields are intended to be within the scope of the following claims.TABLE 1 Poly- Poly- pep- Incyte nucleo- Incyte Incyte tide Poly- tidePoly- Project SEQ ID peptide SEQ ID nucleotide ID NO: ID NO: ID 14161071 1416107CD1 28 1416107CB1 1682513 2 1682513CD1 29 1682513CB1 2446438 32446438CD1 30 2446438CB1 2817822 4 2817822CD1 31 2817822CB1 4009329 54009329CD1 32 4009329CB1 6618083 6 6618083CD1 33 6618083CB1 7472002 77472002CD1 34 7472002CB1 1812692 8 1812692CD1 35 1812692CB1 3232992 93232992CD1 36 3232992CB1 3358383 10 3358383CD1 37 3358383CB1 4250091 114250091CD1 38 4250091CB1 70064803  12 70064803CD1 39 70064803CB170356768  13 70356768CD1 40 70356768CB1 5674114 14 5674114CD1 415674114CB1 1254635 15 1254635CD1 42 1254635CB1 1670595 16 1670595CD1 431670595CB1 1859560 17 1859560CD1 44 1859560CB1 5530164 18 5530164CD1 455530164CB1  139115 19 139115CD1 46 139115CB1 1702940 20 1702940CD1 471702940CB1 1703342 21 1703342CD1 48 1703342CB1 1727529 22 1727529CD1 491727529CB1 2289333 23 2289333CD1 50 2289333CB1 2720354 24 2720354CD1 512720354CB1 3038193 25 3038193CD1 52 3038193CB1 3460979 26 3460979CD1 533460979CB1 7472200 27 7472200CD1 54 7472200CB1

[0365] TABLE 2 Incyte Polypeptide Polypeptide GenBank ProbabilityGenBank SEQ ID NO: ID ID NO: Score Homolog 1 1416107CD1 g7018605 1.9e−302 Glucose transporter [Rattus norvegicus] (Ibberson, M. et al.(2000) J. Biol. Chem. 275:4607-4612) 2 1682513CD1 g5263196  1.4e−153Stretch-inhibitable nonselective channel (SIC) [Rattus norvegicus](Cloning of a stretch-inhibitable nonselective cation channel. J BiolChem. 1999 Mar 5;274 (10) :6330-6335) 3 2446438CD1 g4589141 0 Vanilloidreceptor-like protein 1 [Homo sapiens] (A capsaicin-receptor homologuewith a high threshold for noxious heat. Nature 1999 398:436-441) 54009329CD1 g3873983 1.9e−64 Similar to Na+/Ca+, K+ antiporter [C.elegans] 6 6618083CD1 g9230651  4.7e−268 Facilitative glucosetransporter family member GLUT9 [Homo sapiens] (Phay, J.E. et al. (2000)Genomics 66:217-220) 7 7472002CD1 g433960 0 Aorta CNG channel (rACNG)[Oryctolagus cuniculus] (Primary structure and functional expression ofa cyclic nucleotide- gated channel from rabbit aorta. FEBS Lett. 1993Aug 23;329(1-2) :134-138) 8 1812692CD1 g3928756 4.5e−48 Transientreceptor potential channel 7 [Homo sapiens] (Nagamine, K. et al. (1998)Molecular cloning of a novel putative Ca2+ channel protein (TRPC7)highly expressed in brain) 9 3232992CD1 g3874275 3.2e−70 Similarity toyeast low-affinity glucose transporter HXT4 [Caenorhabditis elegans] 103358383CD1 g3004482  1.4e−163 Putative integral membrane transportprotein [Rattus norvegicus] (Schomig, E. et al. (1998) Molecular cloningand characterization of two novel transport proteins from rat kidney.FEBS Lett. 425:79-86) 11 4250091CD1 g3880445 5.7e−16 VM106R.1 (similarto K+ channel tetramerisation domain) [Caenorhabditis elegans] 1270064803CD1 g3874275 7.0e−84 Similarity to yeast low-affinity glucosetransporter HXT4 [Caenorhabditis elegans] 13 70356768CD1 g183298 4.1e−54GLUT5 protein [Homo sapiens] (Kayano, Y. et al. (1990) Humanfacilitative glucose transporters. Isolation, functionalcharacterization, and gene localization of cDNAs encoding an isoform(GLUT5) expressed in small intestine, kidney, muscle, and adipose tissueand an unusual glucose transporter pseudogene-like sequence (GLUT6). J.Biol. Chem. 265:13276-13282) 14 5674114CD1 g5771352  1.3e−238 Inwardrectifier potassium channel Kir2.4 [Homo sapiens] (Topert, C. et al.(1998) Kir2.4: a novel K+ inward rectifier channel associated withmotoneurons of cranial nerve nuclei. J. Neurosci. 18:4096-4105) 151254635CD1 g3953533   1.76−210 Inwardly rectifying potassium channelKir5.1 [Mus musculus] (Mouri, T. et al. (1998) Assignment of mouseinwardly rectifying potassium channel Kcnj16 to the distal region ofmouse chromosome 11. Genomics 54:181-182) 16 1670595CD1 g9502260 2.3e−146 Cation-chloride cotransporter-interacting protein [Homosapiens] (Caron, L. et al. (2000) J. Biol. Chem. 275:32027- 32036) 171859560CD1 g5834394  1.4e−101 Sulfate transporter [Drosophilamelanogaster] 18 5530164CD1 g4903004 3.5e−20 UDP-N-acetylglucosaminetransporter [Homo sapiens] (Ishida, N. et al. (1999) Molecular cloningand functional expression of the human golgi UDP-N-acetylglucosaminetransporter. J. Biochem. 126:68-77.) 19 139115CD1 g8131858 1.5e−49Putative thymic stromal co-transporter TSCOT [Mus musculus] (Kim, M.G.et al. (2000) J. Immunol. 164:3185-3192) 20 1702940CD1 g5725224 2.5e−143 bK212A2.2 (similar to apolipoprotein L) [Homo sapiens] 211703342CD1 g6003536 8.1e−06 Calcium channel alpha-1 subunit [Bdellouracandida] 22 1727529CD1 g4529890 0.0 NG22 [Homo sapiens] 23 2289333CD1g4539333 5.6e−35 Putative amino acid transport protein [Arabidopsisthaliana] 24 2720354CD1 g3875242 4.3e−38 Similar to mitochrondrialcarrier protein [Caenorhabditis elegans] 26 3460979CD1 g1931644 4.76−08   Membrane protein PTM1 precursor isolog (putative majorfacilitator superfamily transporter) [Arabidopsis thaliana] 277472200CD1 g2811254 2.8e−21 Amiloride-sensitive Na+ channel [Drosophilamelanogaster] (Adams, C.M. et al. (1998) J. Cell Biol. 140:143-152)

[0366] TABLE 3 SEQ Incyte Amino Potential Potential Analytical IDPolypeptide Acid Phosphorylation Glycosylation Signature Sequences,Methods and NO: ID Residues Sites Sites Domains and Motifs Databases 11416107CD1 477 S99 T2 T281 N349 Sugar transporter domain: MOTIFSA29-F474 HMMER-PFAM Sugar transport protein signatures: BLIMPS-V108-L174, L293-S350, G41-I51, BLOCKS L124-V143, Q267-F277, V375-L396,ProfileScan S398-T410 BLIMPS- Glucose transporter signatures: PRINTSF257-Y278, V375-S398, G439-V459 S430 T205 Transmembrane domains: HMMERI259-A279, L293-M313, L320-Y339, Y438-F457 2 1682513CD1 498 S47 T131S286 N278 N411 Glucose transporter signature: MOTIFS P319-N339 BLIMPS-PRINTS T367 S463 T67 N429 Transmembrane domains: HMMER S315 S382A95-Y117, V142-F160, L178-Y201, A200-F219, F244-L263, P319-N339 32446438CD1 764 T64 T329 S23 N570 Ankyrin repeat: MOTIFS R162-C194;F208-S243 HMMER-PFAM Q293-F328 S67 T106 S268 Transmembrane domains:HMMER S339 S348 S353 L386-F405, I463-V486, S464 S468 S667 F538-S557,L623-I642 T692 S697 T720 T101 T115 S325 T414 Y110 Y227 Y333 4 2817822CD1255 T30 S167 T33 Potassium channel signature: MOTIFS T71 S23 T50 R76-T95BLIMPS- S134 T162 T238 PRINTS 5 4009329CD1 584 S258 S321 T70 N60 N125Signal peptide: MOTIFS M1-G29 HMMER Sodium/calcium exchanger proteinHMMER-PFAM domain: I113-Q252, L431-F576 S271 S273 S468 Transmembranedomains: HMMER S514 S62 T132 T101-F121, T166-I189, L234-Y251, Y382-A402,F492-R513, L560-M584 6 6618083CD1 416 T111 S4 S164 N45 N61 N410 Signalpeptide: MOTIFS M1-G37 SPScan Sugar transporter domain: HMMER A30-E416HMMER-PFAM Sugar transport proteins BLIMPS- signatures: BLOCKSA123-L189, S39-V49, ProfileScan I139-M158, Y298-F308 BLIMPS- Glucosetransporter signatures: PRINTS V288-Y309, I356-Q376 S274 T374 S16Transmembrane domains: HMMER S226 S269 V83-V99, I356-L375 7 7472002CD1664 S402 S40 S46 N311 N379 Transmembrane region, cyclic MOTIFSnucleotide gated channel: HMMER-PFAM Y215-I440 BLIMPS- Cyclic nucleotidebinding domain: BLOCKS K469-D565, A460-S476, G478-V501, G516-L525 S93T107 T313 Transmembrane domain: HMMER T337 T381 T422 Y350-I375 S476 S552T591 T606 T634 T2 S35 T124 T208 T418 T448 Y648 8 1812692CD1 242 T95 S35S37 N15 N84 Protein melastatin chromosome BLAST- T124 S204 S221transmembrane PD018035: PRODOM S2 S9 S20 T21 Y117-I227 S86 Y36 93232992CD1 398 T307 S338 N161 Transmembrane domains: HMMER A217-Q242,L247-F264, L350-F368 S100 T133 T241 Sugar transport proteins signature:MOTIFS T303 S377 S395 L45-G94, V30-I96 BLIMPS- BLOCKS ProfileScan 103358383CD1 553 S337 S352 S409 N39 N56 N62 Transmembrane domains: HMMERF204-A222, M470-Y493, I500-T519 T58 S60 S109 N102 N107 glpT family oftransporters: MOTIFS V151-D168 BLIMPS- BLOCKS T133 S337 T433 N473Organic transport protein, renal BLAST- T527 S167 S201 aniontransporter, cationic kidney PRODOM T226 S282 T323 specific solutePD151320: T405 N102-L144 11 4250091CD1 213 S2 S93 S172 N188 Potassiumchannel signature: MOTIFS S184 T17 T22 Q48-T67 BLIMPS- T137 S210 Y89PRINTS 12 70064803CD1 476 T365 S11 S364 Transmembrane domains: HMMERV222-R239, G327-V350, M413-F432 S453 S292 T361 Sugar transport proteinssignature: MOTIFS S390 S466 L153-G202, V138-I204 BLIMPS- BLOCKSProfileScan 13 70356768CD1 246 S100 S238 S118 N34 N50 Signal peptide:M1-G27 MOTIFS Sugar transport proteins signature: SPScan I127-G176,A112-V178, T28-I38, HMMER M128-M147, M133-R158 BLIMPS- BLOCKSProfileScan BLIMPS- PRINTS S215 Transmembrane domain: M163-L181 HMMERSugar transport proteins: BLAST-DOMO DM00135|P46408|: A112-C229 145674114CD1 436 S11 T80 S154 N195 Inward rectifier potassium channelMOTIFS domain: HMMER-PFAM V53-L394, R72-L118, P126-Q169, BLIMPS-PFAMC170-V199, A204-Q238, D296-Y346, T358-E368 S340 S362 T263 Transmembranedomain: W88-L114 HMMER S376 S422 Y47 Inward rectifier potassium channel:BLAST-DOMO DM00448|P52188|: N34-A395 Inward rectifier potassium channel:BLAST- PD001103: V53-Q372 PRODOM KIR2.4 protein: PD124342: A373-P436PD063376: M1-F52 15 1254635CD1 453 S408 T16 T99 Signal peptide: SPScanM1-A32 S416 S29 T209 Transmembrane domains: HMMER F109-A131, V182-I200T216 T250 S375 Inward rectifier potassium channel HMMER-PFAM domain:L72-T399 Inward rectifier potassium ion BLAST - channel subfamilyPD001103: PRODOM K74-K403 Voltage-gated inward rectifier BLAST-potassium channel BIR9, KIR5.1, PRODOM transmembrane PD063375: M36-H73Inward rectifier potassium channel: BLAST-DOMO DM00448|P52185|27-380:K64-L379 Inward rectifier potassium channel BLIMPS-PFAM signature:S91-L137, P145-Q188, S189-R218, A223-R257, N310-C360, A371-W381 161670595CD1 299 S38 T211 S263 N193 N208 Transmembrane domains: HMMERL37-L58, A74-I93, L134-Y154, F159-V179 S33 T187 Sensitive cotransporter,chloride, BLAST-DOMO sodium: DM01178|S06903|1-128: A32-L153 171859560CD1 606 T96 T116 S298 N294 Transmembrane domains: HMMER L45-I63,T396-T421 S571 T572 S595 Sulfate transporter family domain: HMMER-PFAMM137-A468 T245 Sulfate transporters protein BLIMPS- signature: A54-I107,L125-L176 BLOCKS Sulfate transport protein, BLAST- transmembrane,permease PD001255: PRODOM M137-L465 Sulfate transport protein, BLAST-transmembrane, permease PD001121: PRODOM L30-G143 Sulfate transporter:BLAST-DOMO DM01229|S64926|69-531: L30-W428 Sulfate transporters motif:MOTIFS P77-R98 18 5530164CD1 324 S2 S139 Y115 N99 N100 N101Transmembrane domains: HMMER N232 A145-V163, L302-L319 19 139115CD1 445S54 S32 S77 N22 N30 N37 Transmembrane domains: HMMER W182-F200,F242-I261, Y283-F302 S217 S424 S438 N127 N213 Sugar transporter motif:L75-S91 MOTIFS T150 S237 S443 N235 Glucose transporter signature:BLIMPS- Y23 W182-L202 PRINTS 20 1702940CD1 337 T30 S167 T208Apolipoprotein L precursor, lipid BLAST- S306 transport, glycoprotein,signal, PRODOM DJ68O2.1 PD042084: M1-D336 21 1703342CD1 273 T3 S63 T222Transmembrane domains: HMMER F101-A118, M142-F161, F170-I186, I202-I218S248 S250 T10 Ion transport protein domain: HMMER-PFAM S98 S219 S224L95-L269 (Score: −132.1, E-value: 0.72) 22 1727529CD1 710 S31 S102 S119N29 N69 N155 Transmembrane domains: HMMER C38-Y58, V241-L266, W309-V326,F356-T375, F440-L458, T499-I522, L598-F618, I645-V663 T135 S304 S22 N197N298 ABC 3 transport family: S228-Q427 HMMER-PFAM (Score: −182.9,E-value: 2.1) S218 S430 S431 N393 N405 Anion exchanger signature:BLIMPS- T494 S573 S619 N416 N678 A311-L330 PRODOM Y13 23 2289333CD1 476T97 T7 S8 S125 N166 N169 Transmembrane domains: HMMER L54-I81,V127-F145, Y184-S208, L279-G297, I331-K357, I426-T451 T443 S272 S322N212 N425 Transmembrane amino acid HMMER-PFAM transporter proteindomain: A55-F436 T351 T451 Y184 N467 Amino acid transporter protein,BLAST- permease, transmembrane, putative PRODOM proline PD001875:D27-I337 24 2720354CD1 237 T17 T64 S172 Signal peptide: M1-G15 SPScanMitochondrial carrier proteins HMMER-PFAM domain: S25-L109, L122-T202Mitochondrial energy transfer BLIMPS- proteins signature: L128-Q152BLOCKS Mitochondrial energy transfer ProfileScan proteins signature:L27-I75, V123-Q171 Mitochondrial carrier proteins BLIMPS- signature:G87-D107, V136-D154 PRINTS Adenine nucleotide translocator 1 BLIMPS-signature: R63-V84, E176-R191 PRINTS Mitochondrial carrier proteinsMOTIFS motifs: P46-L54, P143-L151 Transport protein, transmembrane,BLAST- inner mitochondrial, ADP/ATP: PRODOM PD000117: L31-F200Mitochondrial energy transfer BLAST-DOMO protein:DM00026|P38087|243-325: L128-Y209 25 3038193CD1 345 T204 T251 S57 N246Transmembrane domains: HMMER L67-L95, I134-I156, I224-F242 S243 T263T308 Sodium bile acid symporter family: HMMER-PFAM Y44-P212 (Score:−7.0, E-value: 9.0e−4) S340 Phosphate transporter signature: BLIMPS-F153-G171 PRODOM 26 3460979CD1 521 S115 T184 S75 N70 N169 N211Transmembrane domains: HMMER L265-L284, I335-I361 T93 S100 S126 Proteinprecursor PTM1, BLAST- S128 S134 S148 transmembrane, signal PD014374:PRODOM S183 S213 S256 G219-E517 S363 S389 S430 (P-value: 7.1e−07) S510T171 S180 S235 T247 S422 Y506 27 7472200CD1 555 T43 S56 T92 N132 N175Amiloride-sensitive sodium channel BLIMPS- alpha subunit signaturePR01078: PRINTS Y102-N118, Y342-Q353, Q353-P370, Q388-N404, G455-E471T148 T298 S423 N311 N361 Transmembrane domain: V452-F475 HMMER S468 S20S52 N421 Amiloride-sensitive sodium channel HMMER-PFAM ASC: F38-L476 S82S96 T184 Amiloride-sensitive sodium channel BLIMPS- S208 S252 S393proteins BL01206: BLOCKS R37-L47, Y342-F368, L427-L472

[0367] TABLE 4 Incyte Polynucleotide Polynucleotide Sequence SelectedSequence 5′ 3′ SEQ ID NO: ID Length Fragments Fragments PositionPosition 28 1416107CB1 2080   1-109, g1941704 116 609  1901-2080,6813453H1 (ADRETUR01) 319 870 1363-1446 6605280H1 (UTREDIT07) 820 1447881845R1 (THYRNOT02) 889 1479 1416107F6 (BRAINOT12) 1348 1896 1416107T6(BRAINOT12) 1579 2080 71826604V1 1563 1974 7448905T1 (BRAYDIN03) 15782050 6300413H1 (UTREDIT07) 423 752 71805807V1 750 1580 71827149V1 15842080 71807187V1 701 1241 6813453R6 (ADRETUR01) 1 312 29 1682513CB1 2128   1-1535, 70207988V1 1 469 1560-1581 70213506V1 394 872 70211216V1 489985 70210573V2 852 1468 70207907V1 988 1512 70210540V2 1357 19482866122T6 (KIDNNOT20) 1548 2108 70211461V1 1597 2128 30 2446438CB1 2825  1-65, 5073532H2 (COLCTUT03) 1 311  2000-2202, 6309494H1 (NERDTDN03)260 812  999-1820 6268005H1 (MCLDTXN03) 344 996 70382927D1 996 152570386205D1 1469 2075 1798255F6 (COLNNOT27) 1632 2194 1562088F6(SPLNNOT04) 2178 2727 2514370F6 (LIVRTUT04) 2303 2825 31 2817822CB1 1718  1-71, 1502510F6 (BRAITUT07) 1 439 609-914 70271734V1 183 76870273052V1 431 930 70271651V1 891 1453 2817822F6 (BRSTNOT14) 981 153870272460V1 1094 1718 32 4009329CB1 2000  1-962 6466193H1 (PLACFEB01) 1640 6780428J1 (OVARDIR01) 582 1260 6307863H1 (NERDTDN03) 725 13646781250H1 (OVARDIR01) 972 1639 7253109J1 (PROSTME05) 1514 1842 6759035J1(HEAONOR01) 1515 2000 33 6618083CB1 2216   1-96, 5722362H1 (SEMVNOT05) 1581 1201-2216 70789558V1 504 1127 70787652V1 588 1203 70791819V1 10501650 70787819V1 1361 1984 70791126V1 1695 2216 34 7472002CB1 1995  1-862, g2121300.v113.gs_2.nt.edit 1 1995 1766-1995 35 1812692CB1 988  1-147, 1812692F6 (PROSTUT12) 564 984 244-570 5425924F6 (PROSTMT07) 1488 g2525933 823 988 5000833F6 (PROSTUT21) 283 804 36 3232992CB1 3179 2106-2665, 224000R6 (PANCNOT01) 2435 3087   1-1646 6825934J1(SINTNOR01) 1 515 7062063H1 (PENITMN02) 2683 3179 4491105H1 (BRAMDIT02)2167 2861 1698347F6 (BLADTUT05) 1762 2333 70053653D1 1392 1870 1807402F6(SINTNOT13) 476 1019 70055908D1 1317 1837 7170824H2 (BRSTTMC01) 719 13736555265H1 (BRAFNON02) 1859 2372 37 3358383CB1 1986  1465-1986, g1444660992 1464 1340-1371 g1009986 768 1254 027195T6 (SPLNFET01) 1674 1986g1505781 552 1252 3358383T6 (PROSTUT16) 1341 1691 6221856U1 585 12526221857U1 1 727 38 4250091CB1 3294   1-920, g715570 2893 3294 1991-2034, 70759966V1 1864 2477  2488-2760, 4250091F6 (BRADDIR01) 1 5321365-1630 5715843H1 (PANCNOT16) 2539 3235 70789723V1 444 1032 966456R6(BRSTNOT05) 2862 3293 70759467V1 1199 1842 70788682V1 604 1302 7056848H1(BRALNON02) 2373 2989 858645R1 (BRAITUT03) 1940 2507 70761829V1 13331947 39 70064803CB1 2043   1-22, 2758549R6 (THP1AZS08) 1540 2043 544-1285 6810024J1 (SKIRNOR01) 576 1280 1676182T6 (BLADNOT05) 1361 20192109762R6 (BRAITUT03) 1181 1797 70503885V1 501 1183 7177480H2(BRAXDIC01) 1 534 40 70356768CB1 1915 1241-1263, 70450108V1 509 1081 853-897, 70451575V1 1072 1730   1-143, 1468307F6 (PANCTUT02) 1 524 1450-1532, 70451567V1 368 1078 668-820 70449058V1 1384 1915 70449392V11060 1720 41 5674114CB1 1809  1-402 6776218J1 (OVARDIR01) 1078 18093024042H1 (PROSDIN01) 690 1040 6292787H1 (BMARUNA01) 939 1290 6776218H1(OVARDIR01) 184 944 g5686663.v113.gs_16.nt 1 1311 42 1254635CB1 1730  1-106, 2613664F6 (ESOGTUT02) 655 1177  1711-1730, SXBC01035V1 75 573 567-635, 2863343F6 (KIDNNOT20) 1 515 696-900 2614317T6 (GBLANOT01) 11261730 SCSA01493V1 548 724 3323244T6 (PTHYNOT03) 960 1627 43 1670595CB11147  746-980, SCIA02891V1 368 1147  1-696 SCIA04658V1 1 641 441859560CB1 2745   1-820, 6195927H1 (PITUNON01) 2367 2745  865-2059824186R1 (PROSNOT06) 1159 1718 1399644F6 (BRAITUT08) 503 973 6812696J1(ADRETUR01) 30 732 7255024H1 (FIBRTXC01) 814 1364 2127239R7 (KIDNNOT05)1732 2176 5990868H1 (FTUBTUT02) 1493 1795 6826978H1 (SINTNOR01) 1 4796850265H1 (BRAIFEN08) 2056 2740 4677711H1 (NOSEDIT02) 1107 13811859560T6 (PROSNOT18) 2247 2745 4320381H1 (BRADDIT02) 1925 2202 455530164CB1 3204  1241-2064, 7175713H1 (BRSTTMC01) 178 811   1-548,3217236H1 (TESTNOT07) 530 812  572-1097 2552002H1 (LUNGTUT06) 1 24270039789V1 2452 3175 70090155V1 1781 2492 6830248J1 (SINTNOR01) 542 12116729730H1 (COLITUT02) 1214 1889 2850920F6 (BRSTTUT13) 929 1416 6125934H1(BRAHNON05) 2505 3204 6059181H1 (BRAENOT04) 1923 2496 1956752F6(CONNNOT01) 1452 1893 46 139115CB1 2763   1-770, 6455739H1 (COLNDIC01)528 1219  1345-1389, 70077060U1 1900 2535 1455-1683 7126228H1(COLNDIY01) 1 580 2468105F6 (THYRNOT08) 827 1463 70079483U1 2145 276370122236V1 1424 1971 70122363V1 1451 1976 351595R1 (LVENNOT01) 2066 256647 1702940CB1 1639   1-246, 70480521V1 1027 1639 1536-1639 4335403F6(KIDCTMT01) 1 516 70466195V1 559 1157 70466476V1 494 1102 48 1703342CB11600  1-812 3348562H1 (BRAITUT24) 1 282 285125R1 (EOSIHET02) 1041 15987071066H1 (BRAUTDR02) 250 854 6494627H1 (BONRNOT01) 1220 1600 6879086J1(LNODNOR03) 360 1094 49 1727529CB1 2380   1-569, 957891H1 (KIDNNOT05)2085 2380 1228-1654 60211961U1 691 1237 6800135J1 (COLENOR03) 1250 19836798918J1 (COLENOR03) 1693 2284 60211964U1 262 807 6798894H1 (COLENOR03)1089 1774 3249035F6 (SEMVNOT03) 1 626 3566495H1 (BRONNOT02) 1984 2298 502289333CB1 3038   1-611, 2552315T6 (LUNGTUT06) 1322 1862 2497-2524g872898 848 1328 1435329F1 (PANCNOT08) 2197 2725 3553901H1 (SYNONOT01)2570 2865 2508452H1 (CONUTUT01) 1 114 2771704H1 (COLANOT02) 1815 20786999443H1 (HEALDIR01) 2 553 g1665184 2594 3038 2289333R6 (BRAINON01)1254 1708 g1156003 2524 3032 5597992H1 (UTRENON03) 1029 1286 g5545742612 1066 5836345H1 (BRAIDIT05) 2624 2880 4220788F6 (PANCNOT07) 613 9581994713T6 (BRSTTUT03) 1949 2451 2040880R6 (HIPONON02) 463 821 512720354CB1 2608   1-2058 2720354F6 (LUNGTUT10) 490 1046 6942433H1(FTUBTUR01) 885 1437 6121303H1 (BRAHNON05) 1630 2340 6558224H1(BRAFNON02) 1713 2406 6940932H1 (FTUBTUR01) 1 465 g1927466 325 8726826181J1 (SINTNOR01) 1062 1666 6197805H1 (PITUNON01) 2154 2608 523038193CB1 3804  3392-3457, 044564H1 (TBLYNOT01) 2311 2562  1169-1264,901446R6 (BRSTTUT03) 1733 2282   1-829, g4088232 3459 3804  2271-2483,1428831H1 (SINTBST01) 473 661 1319-1363 2741328T6 (BRSTTUT14) 3235 38044970206H1 (KIDEUNC10) 1029 1303 2768967H1 (COLANOT02) 772 1020 5688762F6(BRAIUNT01) 36 621 3038193F6 (BRSTNOT16) 1284 1710 6477440H1 (PROSTMC01)1843 2486 70809191V1 2411 2832 3154867H1 (TLYMTXT02) 1 272 2257401R6(OVARTUT01) 2660 3167 g4268882 1421 1815 2257401T6 (OVARTUT01) 2896 352053 3460979CB1 1894  1-36 2237852F6 (PANCTUT02) 519 945 1746-18943460979F6 (293TF1T01) 1000 1500 7161336H1 (PLACNOR01) 582 1193 6800921J1(COLENOR03) 1 557 7057496H1 (BRALNON02) 1206 1894 54 7472200CB1 1668  1-1668 GNN.g6554406_006 1 1668

[0368] TABLE 5 Polynucleotide Incyte Representative SEQ ID NO: ProjectID Library 28 1416107CB1 UTREDIT07 29 1682513CB1 SPLNNOT11 30 2446438CB1MCLDTXN03 31 2817822CB1 BRAITUT07 32 4009329CB1 OVARDIN02 33 6618083CB1HELAUNT01 35 1812692CB1 PROSTUT12 36 3232992CB1 PANCNOT01 37 3358383CB1PROSTUT16 38 4250091CB1 BRAITUT03 39 70064803CB1 THP1AZS08 4070356768CB1 HNT2AGT01 41 5674114CB1 OVARDIR01 42 1254635CB1 LUNGFET03 431670595CB1 BRAITUT24 44 1859560CB1 NGANNOT01 45 5530164CB1 BRAYDIN03 46139115CB1 SINTNOT18 47 1702940CB1 BRAVTXT04 48 1703342CB1 EOSIHET02 491727529CB1 PROSNOT18 50 2289333CB1 LUNGTUT06 51 2720354CB1 PROSTUS23 523038193CB1 LIVRNON08 53 3460979CB1 COLENOR03

[0369] TABLE 6 Library Vector Library Description UTREDIT07 pINCYLibrary was constructed using RNA isolated from diseased endometrialtissue removed from a female during endometrial biopsy. Pathologyindicated in phase endometrium with missing beta 3, Type II defects.SPLNNOT11 pINCY Library was constructed using RNA isolated from diseasedspleen tissue removed from a 14- year-old Asian male during a totalsplenectomy. Pathology indicated changes consistent with idopathicthrombocytopenic purpura. The patient presented with bruising. Patientmedications included Vincristine. MCLDTXN03 pINCY Library wasconstructed from a pool of two dendritic cell libraries. Startinglibraries were constructed using RNA isolated from untreated and treatedderived dendritic cells from umbilical cord blood CD34+ precursor cellsremoved from a male. The cells were derived with granulocyte/macrophagecolony stimulating factor (GM-CSF), tumor necrosis factor alpha (TNFalpha), and stem cell factor (SCF). The libraries were normalized underconditions adapted from Soares et al. (1994) Proc. Natl. Acad. Sci. USA91:9228 and Bonaldo et al. (1996) Genome Res. 6:791, except that asignificantly longer (48 hours/round) reannealing hybridization wasused. BRAITUT07 pINCY Library was constructed using RNA isolated fromleft frontal lobe tumor tissue removed from the brain of a 32-year-oldCaucasian male during excision of a cerebral meningeal lesion. Pathologyindicated low grade desmoplastic neuronal neoplasm. The patientpresented with nausea, vomiting, and headache. Patient history includedalcohol, tobacco use, and marijuana use twice a week for six years.Family history included atherosclerotic coronary artery disease in thegrandparent(s). OVARDIN02 pINCY Library was constructed from an ovariantissue library. Starting RNA was made from diseased ovarian tissueremoved from a 39-year-old Caucasian female during total abdominalhysterectomy, bilateral salpingo-oophorectomy, dilation and curettage,partial colectomy, incidental appendectomy, and temporary colostomy.Pathology indicated the right and left adnexa, mesentery and muscularispropria of the sigmoid colon were extensively involved by endometriosis.Endometriosis also involved the anterior and posterior serosal surfacesof the uterus and the cul-de-sac. The endometrium was proliferative.Pathology for the associated tumor tissue indicated multiple (3intramural, 1 subserosal) leiomyomata. The patient presented withabdominal pain and infertility. Patient history included scoliosis.Previous surgeries included laparoscopic cholecystectomy and exploratorylaparotomy. Patient medications included Megace, Danazol, and Lupron.Family history included hyperlipidemia in the mother, benignhypertension, hyperlipidemia, atherosclerotic coronary artery disease,coronary artery bypass graft, depressive disorder, brain cancer, andtype II diabetes. The library was normalized under conditions adaptedfrom Soares et al. (1994) Proc. Natl. Acad. Sci. USA 91:9228 and Bonaldoet al. (1996) Genome Res. 6:791, except that a significantly longer (48hours/round) reannealing hybridization was used. HELAUNT01 pINCY Librarywas constructed from RNA isolated from an untreated HeLa cell line,derived from cervical adenocarcinoma removed from a 31-year-old Blackfemale. BRAITUT03 PSPORT1 Library was constructed using RNA isolatedfrom brain tumor tissue removed from the left frontal lobe of a17-year-old Caucasian female during excision of a cerebral meningeallesion. Pathology indicated a grade 4 fibrillary giant and small-cellastrocytoma. Family history included benign hypertension andcerebrovascular disease. HNT2AGT01 PBLUESCRIPT Library was constructedat Stratagene (STR937233), using RNA isolated from the hNT2 cell linederived from a human teratocarcinoma that exhibited propertiescharacteristic of a committed neuronal precursor. Cells were treatedwith retinoic acid for 5 weeks and with mitotic inhibitors for two weeksand allowed to mature for an additional 4 weeks in conditioned medium.OVARDIR01 pcDNA2.1 Library was constructed using RNA isolated from rightovary tissue removed from a 45- year-old Caucasian female during totalabdominal hysterectomy, bilateral salpingo- oophorectomy, vaginalsuspension and fixation, and incidental appendectomy. Pathologyindicated stromal hyperthecosis of the right and left ovaries. Pathologyfor the matched tumor tissue indicated a dermoid cyst (benign cysticteratoma) in the left ovary. Multiple (3) intramural leiomyomata wereidentified. The cervix showed squamous metaplasia. Patient historyincluded metrorrhagia, female stress incontinence, alopecia, depressivedisorder, pneumonia, normal delivery, and deficiency anemia. Familyhistory included benign hypertension, atherosclerotic coronary arterydisease, hyperlipidemia, and primary tuberculous complex. PANCNOT01PBLUESCRIPT Library was constructed using RNA isolated from thepancreatic tissue of a 29-year- old Caucasian male who died from headtrauma. PROSTUT12 pINCY Library was constructed using RNA isolated fromprostate tumor tissue removed from a 65-year-old Caucasian male during aradical prostatectomy. Pathology indicated an adenocarcinoma (Gleasongrade 2 + 2). Adenofibromatous hyperplasia was also present. The patientpresented with elevated prostate specific antigen (PSA). PROSTUT16 pINCYLibrary was constructed using RNA isolated from prostate tumor tissueremoved from a 55-year-old Caucasian male. Pathology indicatedadenocarcinoma, Gleason grade 5 + 4. Adenofibromatous hyperplasia wasalso present. The patient presented with elevated prostate specificantigen (PSA). Patient history included calculus of the kidney. Familyhistory included lung cancer and breast cancer. THP1AZS08 PSPORT1 Thissubtracted THP-1 promonocyte cell line library was constructed using5.76 × 1e6 clones from a 5-aza-2′-deoxycytidine (AZ) treated THP-1 celllibrary. Starting RNA was made from THP-1 promonocyte cells treated forthree days with 0.8 micromolar AZ. The hybridization probe forsubtraction was derived from a similarly constructed library, made fromRNA isolated from untreated THP-1 cells. 5.76 million clones from theAZ-treated THP-1 cell library were then subjected to two rounds ofsubtractive hybridization with 5 million clones from the untreated THP-1cell library. Subtractive hybridization conditions were based on themethodologies of Swaroop et al., NAR (1991) 19:1954, and Bonaldo et al.,Genome Research (1996) 6:791. THP-1 (ATCC TIB 202) is a humanpromonocyte line derived from peripheral blood of a 1-year- oldCaucasian male with acute monocytic leukemia (ref: Int. J. Cancer 26(1980) :171). BRAITUT24 pINCY Library was constructed using RNA isolatedfrom right frontal brain tumor tissue removed from a 50-year-oldCaucasian male during a cerebral meninges lesion excision. Pathologyindicated meningioma. Family history included colon cancer andcerebrovascular disease. BRAYDIN03 pINCY This normalized brain tissuelibrary was constructed from 6.7 million independent clones from a braintissue library. Starting RNA was made from RNA isolated from diseasedhypothalamus tissue removed from a 57-year-old Caucasian male who diedfrom a cerebrovascular accident. Patient history included Huntington'sdisease and emphysema. The library was normalized in 2 rounds usingconditions adapted from Scares et al., PNAS (1994) 91:9228 and Bonaldoet al., Genome Research 6 (1996) ;791, except that a significantlylonger (48 hours/round) reannealing hybridization was used. The librarywas linearized and recircularized to select for insert containingclones. LUNGFET03 pINCY Library was constructed using RNA isolated fromlung tissue removed from a Caucasian female fetus, who died at 20 weeks'gestation. NGANNOT01 PSPORT1 Library was constructed using RNA isolatedfrom tumorous neuroganglion tissue removed from a 9-year-old Caucasianmale during a soft tissue excision of the chest wall. Pathologyindicated a ganglioneuroma. Family history included asthma. BRAVTXT04PSPORT1 Library was constructed using RNA isolated from separatepopulations of human astrocytes stimulated for 4 to 6 hours with acombination of cytokines including IL- 1. The RNA was pooled for polyARNA isolation and library construction. EOSIHET02 PBLUESCRIPT Librarywas constructed using RNA isolated from peripheral blood cells apheresedfrom a 48-year-old Caucasian male. Patient history includedhypereosinophilia. The cell population was determined to be greater than77% eosinophils by Wright's staining. LIVRNON08 pINCY This normalizedliver tissue library was constructed from 5.7 million independent clonesfrom a pooled liver tissue library. Starting RNA was isolated frompooled liver tissue removed from a 4-year-old Hispanic male who diedfrom anoxia and a 16 week female fetus who died after 16-weeks gestationfrom anencephaly. Serologies were positive for cytolomegalovirus in the4-year-old. Patient history included asthma in the 4-year-old. Familyhistory included taking daily prenatal vitamins and mitral valveprolapse in the mother of the fetus. The library was normalized in 2rounds using conditions adapted from Scares et al. Proc. Natl. Acad.Sci. USA (1994) 91:9228 and Bonaldo et al. (1996) Genome Research 6:791,except that a significantly longer (48 hours/round) reannealinghybridization was used. LUNGTUT06 pINCY Library was constructed usingRNA isolated from apical lung tumor tissue removed from an 80-year-oldCaucasian female during a segmental lung resection. Pathology indicateda metastatic granulosa cell tumor. Patient history included pelvic softtissue tumor and chemotherapy for one year. Family history includedtuberculosis, lung cancer, and atherosclerotic coronary artery disease.PROSNOT18 pINCY Library was constructed using RNA isolated from diseasedprostate tissue removed from a 58-year-old Caucasian male during aradical cystectomy, radical prostatectomy, and gastrostomy. Pathologyindicated adenofibromatous hyperplasia; this tissue was associated witha grade 3 transitional cell carcinoma. Patient history included anginaand emphysema. Family history included acute myocardial infarction,atherosclerotic coronary artery disease, and type II diabetes. PROSTUS23pINCY This subtracted prostate tumor library was constructed using 1million clones from a pooled prostate tumor library that was subjectedto 2 rounds of subtractive hybridization with 1 million clones from apooled prostate tissue library. The starting library for subtraction wasconstructed by pooling equal numbers of clones from 4 prostate tumorlibraries using mRNA isolated from prostate tumor removed from Caucasianmales at ages 58 (A), 61 (B), 66 (C), and 68 (D) during prostatectomywith lymph node excision. Pathology indicated adenoCA in all donors.History included elevated PSA, induration and tobacco abuse in donor A;elevated PSA, induration, prostate hyperplasia, renal failure,osteoarthritis, renal artery stenosis, benign HTN, thrombocytopenia,hyperlipidemia, tobacco/alcohol and hepatitis C (carrier) in donor B;elevated PSA, induration, and tobacco abuse in donor C; and elevatedPSA, induration, hypercholesterolemia, and kidney calculus in donor D.The hybridization probe for subtraction was constructed by pooling equalnumbers of cDNA clones from 3 prostate tissue libraries derived fromprostate tissue, prostate epithelial cells, and fibroblasts fromprostate stroma from 3 different donors. Subtractive hybridizationconditions were based on the methodologies of Swaroop et al. (1991)Nucleic Acids Res. 19:1954 and Bonaldo et al. Genome Research (1996)6:791. SINTNOT18 pINCY Library was constructed using RNA isolated fromsmall intestine tissue obtained from a 59-year-old male. COLENOR03PCDNA2.1 Library was constructed using RNA isolated from colonepithelium tissue removed from a 13-year-old Caucasian female who diedfrom a motor vehicle accident.

[0370] TABLE 7 Program Description Reference Parameter Threshold ABI Aprogram that removes Applied Biosystems, Foster City, CA. FACTURA vectorsequences and masks ambiguous bases in nucleic acid sequences. ABI/ AFast Data Finder useful Applied Biosystems, Foster City, CA; Mismatch<50% PARACEL in comparing and Paracel Inc., Pasadena, CA. FDF annotatingamino acid or nucleic acid sequences. ABI A program that assemblesApplied Biosystems, Foster City, CA. Auto- nucleic acid sequences.Assembler BLAST A Basic Local Alignment Altschul, S. F. et al. (1990) J.Mol. Biol. ESTs: Probability Search Tool useful in 215:403-410;Altschul, S. F. et al. (1997) value = 1.0E−8 sequence similarity searchNucleic Acids Res. 25:3389-3402. or less for amino acid and Full Lengthsequences: nucleic acid sequences. Probability BLAST includes five value= 1.0E−10 functions: blastp, blastn, or less blastx, tblastn, andtblastx. FASTA A Pearson and Lipman Pearson, W. R. and D. J. Lipman(1988) Proc. ESTs: fasta E value = algorithm that searches for Natl.Acad Sci. USA 85:2444-2448; Pearson, 1.06E−6 similarity between a queryW. R. (1990) Methods Enzymol. 183:63-98; Assembled ESTs: fasta sequenceand a group of and Smith, T. F. and M. S. Waterman (1981) Identity =sequences of the same type. Adv. Appl. Math. 2:482-489. 95% or greaterand FASTA comprises as Match length = 200 least five functions: fasta,bases or greater; tfasta, fastx, tfastx, and fastx E value = ssearch.1.0E−8 or less Full Length sequences: fastx score = 100 or greaterBLIMPS A BLocks IMProved Searcher Henikoff, S. and J. G. Henikoff (1991)Nucleic Probability value = that matches a Acids Res. 19:6565-6572;Henikoff, J. G. and 1.0E−3 or less sequence against those in S. Henikoff(1996) Methods Enzymol. BLOCKS, PRINTS, 266:88-105; and Attwood, T. K.et al. (1997) J. DOMO, PRODOM, and PFAM Chem. Inf. Comput. Sci.37:417-424. databases to search for gene families, sequence homology,and structural fingerprint regions. HMMER An algorithm for searchingKrogh, A. et al. (1994) J. Mol. Biol. PFAM hits: Probability a querysequence against 235:1501-1531; Sonnhammer, E. L. L. et al. value =hidden Markov model (HMM)- (1988) Nucleic Acids Res. 26:320-322; 1.0E−3or less based databases of Durbin, R. et al. (1998) Our World View, in aSignal peptide hits: protein family consensus Nutshell, Cambridge Univ.Press, pp. 1-350. Score = 0 or sequences, such as PFAM. greater Profile-An algorithm that searches Gribskov, M. et al. (1988) CABIOS 4:61-66;Normalized quality Scan for structural and sequence Gribskov, M. et al.(1989) Methods Enzymol. score ≧ GCG- motifs in protein sequences183:146-159; Bairoch, A. et al. (1997) specified “HIGH” that matchsequence patterns Nucleic Acids Res. 25:217-221. value for that definedin Prosite. particular Prosite motif. Generally, score = 1.4-2.1. PhredA base-calling algorithm that Ewing, B. et al. (1998) Genome Res.examines automated 8:175-185; Ewing, B. and P. Green sequencer traceswith high (1998) Genome Res. 8:186-194. sensitivity and probability.Phrap A Phils Revised Assembly Smith, T. F. and M. S. Waterman (1981)Adv. Score = 120 Program including SWAT and Appl. Math. 2:482-489;Smith, T. F. and M. S. or greater; CrossMatch, programs based Waterman(1981) J. Mol. Biol. 147:195-197; Match on efficient implementation andGreen, P., University of Washington, length = of the Smith-WatermanSeattle, WA. 56 or greater algorithm, useful in searching sequencehomology and assembling DNA sequences. Consed A graphical tool forviewing Gordon, D. et al. (1998) Genome Res. 8:195-202. and editingPhrap assemblies. SPScan A weight matrix analysis Nielson, H. et al.(1997) Protein Engineering Score = 3.5 program that scans protein10:1-6; Claverie, J. M. and S. Audic (1997) or greater sequences for thepresence of CABIOS 12:431-439. secretory signal peptides. TMAP A programthat uses weight Persson, B. and P. Argos (1994) J. Mol. Biol. matricesto delineate 237:182-192; Persson, B. and P. Argos (1996) transmembranesegments on Protein Sci. 5:363-371. protein sequences and determineorientation. TMHMMER A program that uses a hidden Sonnhammer, E. L. etal. (1998) Proc. Sixth Intl. Markov model (HMM) to Conf. on IntelligentSystems for Mol. Biol., delineate transmembrane Glasgow et al., eds.,The Am. Assoc. for Artificial segments on protein sequences IntelligencePress, Menlo Park, CA, pp. 175-182. and determine orientation. Motifs Aprogram that searches amino Bairoch, A. et al. (1997) Nucleic Acids Res.25:217-221; acid sequences for patterns Wisconsin Package ProgramManual, version 9, page that matched those defined M51-59, GeneticsComputer Group, Madison, WI. in Prosite.

[0371]

1 54 1 477 PRT Homo sapiens misc_feature Incyte ID No 1416107CD1 1 MetThr Pro Glu Asp Pro Glu Glu Thr Gln Pro Leu Leu Gly Pro 1 5 10 15 ProGly Gly Ser Ala Pro Arg Gly Arg Arg Val Phe Leu Ala Ala 20 25 30 Phe AlaAla Ala Leu Gly Pro Leu Ser Phe Gly Phe Ala Leu Gly 35 40 45 Tyr Ser SerPro Ala Ile Pro Ser Leu Gln Arg Ala Ala Pro Pro 50 55 60 Ala Pro Arg LeuAsp Asp Ala Ala Ala Ser Trp Phe Gly Ala Val 65 70 75 Val Thr Leu Gly AlaAla Ala Gly Gly Val Leu Gly Gly Trp Leu 80 85 90 Val Asp Arg Ala Gly ArgLys Leu Ser Leu Leu Leu Cys Ser Val 95 100 105 Pro Phe Val Ala Gly PheAla Val Ile Thr Ala Ala Gln Asp Val 110 115 120 Trp Met Leu Leu Gly GlyArg Leu Leu Thr Gly Leu Ala Cys Gly 125 130 135 Val Ala Ser Leu Val AlaPro Val Tyr Ile Ser Glu Ile Ala Tyr 140 145 150 Pro Ala Val Arg Gly LeuLeu Gly Ser Cys Val Gln Leu Met Val 155 160 165 Val Val Gly Ile Leu LeuAla Tyr Leu Ala Gly Trp Val Leu Glu 170 175 180 Trp Arg Trp Leu Ala ValLeu Gly Cys Val Pro Pro Ser Leu Met 185 190 195 Leu Leu Leu Met Cys PheMet Pro Glu Thr Pro Arg Phe Leu Leu 200 205 210 Thr Gln His Arg Arg GlnGlu Ala Met Ala Ala Leu Arg Phe Leu 215 220 225 Trp Gly Ser Glu Gln GlyTrp Glu Asp Pro Pro Ile Gly Ala Glu 230 235 240 Gln Ser Phe His Leu AlaLeu Leu Arg Gln Pro Gly Ile Tyr Lys 245 250 255 Pro Phe Ile Ile Gly ValSer Leu Met Ala Phe Gln Gln Leu Ser 260 265 270 Gly Val Asn Ala Val MetPhe Tyr Ala Glu Thr Ile Phe Glu Glu 275 280 285 Ala Lys Phe Lys Asp SerSer Leu Ala Ser Val Val Val Gly Val 290 295 300 Ile Gln Val Leu Phe ThrAla Val Ala Ala Leu Ile Met Asp Arg 305 310 315 Ala Gly Arg Arg Leu LeuLeu Val Leu Ser Gly Val Val Met Val 320 325 330 Phe Ser Thr Ser Ala PheGly Ala Tyr Phe Lys Leu Thr Gln Gly 335 340 345 Gly Pro Gly Asn Ser SerHis Val Ala Ile Ser Ala Pro Val Ser 350 355 360 Ala Gln Pro Val Asp AlaSer Val Gly Leu Ala Trp Leu Ala Val 365 370 375 Gly Ser Met Cys Leu PheIle Ala Gly Phe Ala Val Gly Trp Gly 380 385 390 Pro Ile Pro Trp Leu LeuMet Ser Glu Ile Phe Pro Leu His Val 395 400 405 Lys Gly Val Ala Thr GlyIle Cys Val Leu Thr Asn Trp Leu Met 410 415 420 Ala Phe Leu Val Thr LysGlu Phe Ser Ser Leu Met Glu Val Leu 425 430 435 Arg Pro Tyr Gly Ala PheTrp Leu Ala Ser Ala Phe Cys Ile Phe 440 445 450 Ser Val Leu Phe Thr LeuPhe Cys Val Pro Glu Thr Lys Gly Lys 455 460 465 Thr Leu Glu Gln Ile ThrAla His Phe Glu Gly Arg 470 475 2 498 PRT Homo sapiens misc_featureIncyte ID No 1682513CD1 2 Met Arg Arg Gln Asp Ser Arg Gly Asn Thr ValLeu His Ala Leu 1 5 10 15 Val Ala Ile Ala Asp Asn Thr Arg Glu Asn ThrLys Phe Val Thr 20 25 30 Lys Met Tyr Asp Leu Leu Leu Leu Lys Cys Ala ArgLeu Phe Pro 35 40 45 Asp Ser Asn Leu Glu Ala Val Leu Asn Asn Asp Gly LeuSer Pro 50 55 60 Leu Met Met Ala Ala Lys Thr Gly Lys Ile Gly Asn Arg HisGlu 65 70 75 Met Leu Ala Val Glu Pro Ile Asn Glu Leu Leu Arg Asp Lys Trp80 85 90 Arg Lys Phe Gly Ala Val Ser Phe Tyr Ile Asn Val Val Ser Tyr 95100 105 Leu Cys Ala Met Val Ile Phe Thr Leu Thr Ala Tyr Tyr Gln Pro 110115 120 Leu Glu Gly Thr Pro Pro Tyr Pro Tyr Arg Thr Thr Val Asp Tyr 125130 135 Leu Arg Leu Ala Gly Glu Val Ile Thr Leu Phe Thr Gly Val Leu 140145 150 Phe Phe Phe Thr Asn Ile Lys Asp Leu Phe Met Lys Lys Cys Pro 155160 165 Gly Val Asn Ser Leu Phe Ile Asp Gly Ser Phe Gln Leu Leu Tyr 170175 180 Phe Ile Tyr Ser Val Leu Val Ile Val Ser Ala Ala Leu Tyr Leu 185190 195 Ala Gly Ile Glu Ala Tyr Leu Ala Val Met Val Phe Ala Leu Val 200205 210 Leu Gly Trp Met Asn Ala Leu Tyr Phe Thr Arg Gly Leu Lys Leu 215220 225 Thr Gly Thr Tyr Ser Ile Met Ile Gln Lys Ile Leu Phe Lys Asp 230235 240 Leu Phe Arg Phe Leu Leu Val Tyr Leu Leu Phe Met Ile Gly Tyr 245250 255 Ala Ser Ala Leu Val Ser Leu Leu Asn Pro Cys Ala Asn Met Lys 260265 270 Val Cys Asn Gly Asp Gln Thr Asn Cys Thr Val Pro Thr Tyr Pro 275280 285 Ser Cys Arg Asp Ser Glu Thr Phe Ser Thr Phe Leu Leu Asp Leu 290295 300 Phe Lys Leu Thr Ile Gly Met Gly Asp Leu Glu Met Leu Ser Ser 305310 315 Thr Lys Tyr Pro Val Val Phe Ile Ile Leu Leu Val Thr Tyr Ile 320325 330 Ile Leu Thr Phe Val Leu Leu Leu Asn Met Leu Ile Ala Leu Met 335340 345 Gly Glu Thr Val Gly Gln Val Ser Lys Glu Ser Lys His Ile Trp 350355 360 Lys Leu Gln Trp Ala Thr Thr Ile Leu Asp Ile Glu Arg Ser Phe 365370 375 Pro Val Phe Leu Arg Lys Ser Phe Arg Ser Gly Glu Met Val Thr 380385 390 Val Gly Lys Ser Ser Asp Gly Thr Pro Asp Arg Arg Trp Cys Phe 395400 405 Arg Val Asp Glu Val Asn Trp Ser His Trp Asn Gln Asn Leu Gly 410415 420 Ile Ile Asn Glu Asp Pro Gly Lys Asn Glu Thr Tyr Gln Tyr Tyr 425430 435 Gly Phe Ser His Thr Val Gly Arg Leu Arg Arg Asp Arg Trp Ser 440445 450 Ser Val Val Pro Arg Val Val Glu Leu Asn Lys Asn Ser Asn Pro 455460 465 Asp Glu Val Val Val Pro Leu Asp Ser Thr Gly Asn Pro Arg Cys 470475 480 Asp Gly His Gln Gln Gly Tyr Pro Arg Lys Trp Arg Thr Asp Asp 485490 495 Ala Pro Leu 3 764 PRT Homo sapiens misc_feature Incyte ID No2446438CD1 3 Met Thr Ser Pro Ser Ser Ser Pro Val Phe Arg Leu Glu Thr Leu1 5 10 15 Asp Ala Gly Gln Glu Asp Gly Ser Glu Ala Asp Arg Gly Lys Leu 2025 30 Asp Phe Gly Ser Gly Leu Pro Pro Met Glu Ser Gln Phe Gln Gly 35 4045 Glu Asp Arg Lys Phe Ala Pro Gln Ile Arg Val Asn Leu Asn Tyr 50 55 60Arg Lys Gly Thr Gly Ala Ser Gln Pro Asp Pro Asn Arg Phe Asp 65 70 75 ArgAsp Arg Leu Phe Asn Ala Val Ser Arg Gly Val Pro Glu Asp 80 85 90 Leu AlaGly Leu Pro Glu Tyr Leu Ser Lys Thr Ser Lys Tyr Leu 95 100 105 Thr AspSer Glu Tyr Thr Glu Gly Ser Thr Gly Lys Thr Cys Leu 110 115 120 Met LysAla Val Leu Asn Leu Lys Asp Gly Val Asn Ala Cys Ile 125 130 135 Leu ProLeu Leu Gln Ile Asp Arg Asp Ser Gly Asn Pro Gln Pro 140 145 150 Leu ValAsn Ala Gln Cys Thr Asp Asp Tyr Tyr Arg Gly His Ser 155 160 165 Ala LeuHis Ile Ala Ile Glu Lys Arg Ser Leu Gln Cys Val Lys 170 175 180 Leu LeuVal Glu Asn Gly Ala Asn Val His Ala Arg Ala Cys Gly 185 190 195 Arg PhePhe Gln Lys Gly Gln Gly Thr Cys Phe Tyr Phe Gly Glu 200 205 210 Leu ProLeu Ser Leu Ala Ala Cys Thr Lys Gln Trp Asp Val Val 215 220 225 Ser TyrLeu Leu Glu Asn Pro His Gln Pro Ala Ser Leu Gln Ala 230 235 240 Thr AspSer Gln Gly Asn Thr Val Leu His Ala Leu Val Met Ile 245 250 255 Ser AspAsn Ser Ala Glu Asn Ile Ala Leu Val Thr Ser Met Tyr 260 265 270 Asp GlyLeu Leu Gln Ala Gly Ala Arg Leu Cys Pro Thr Val Gln 275 280 285 Leu GluAsp Ile Arg Asn Leu Gln Asp Leu Thr Pro Leu Lys Leu 290 295 300 Ala AlaLys Glu Gly Lys Ile Glu Ile Phe Arg His Ile Leu Gln 305 310 315 Arg GluPhe Ser Gly Leu Ser His Leu Ser Arg Lys Phe Thr Glu 320 325 330 Trp CysTyr Gly Pro Val Arg Val Ser Leu Tyr Asp Leu Ala Ser 335 340 345 Val AspSer Cys Glu Glu Asn Ser Val Leu Glu Ile Ile Ala Phe 350 355 360 His CysLys Ser Pro His Arg His Arg Met Val Val Leu Glu Pro 365 370 375 Leu AsnLys Leu Leu Gln Ala Lys Trp Asp Leu Leu Ile Pro Lys 380 385 390 Phe PheLeu Asn Phe Leu Cys Asn Leu Ile Tyr Met Phe Ile Phe 395 400 405 Thr AlaVal Ala Tyr His Gln Pro Thr Leu Lys Lys Gln Ala Ala 410 415 420 Pro HisLeu Lys Ala Glu Val Gly Asn Ser Met Leu Leu Thr Gly 425 430 435 His IleLeu Ile Leu Leu Gly Gly Ile Tyr Leu Leu Val Gly Gln 440 445 450 Leu TrpTyr Phe Trp Arg Arg His Val Phe Ile Trp Ile Ser Phe 455 460 465 Ile AspSer Tyr Phe Glu Ile Leu Phe Leu Phe Gln Ala Leu Leu 470 475 480 Thr ValVal Ser Gln Val Leu Cys Phe Leu Ala Ile Glu Trp Tyr 485 490 495 Leu ProLeu Leu Val Ser Ala Leu Val Leu Gly Trp Leu Asn Leu 500 505 510 Leu TyrTyr Thr Arg Gly Phe Gln His Thr Gly Ile Tyr Ser Val 515 520 525 Met IleGln Lys Val Ile Leu Arg Asp Leu Leu Arg Phe Leu Leu 530 535 540 Ile TyrLeu Val Phe Leu Phe Gly Phe Ala Val Ala Leu Val Ser 545 550 555 Leu SerGln Glu Ala Trp Arg Pro Glu Ala Pro Thr Gly Pro Asn 560 565 570 Ala ThrGlu Ser Val Gln Pro Met Glu Gly Gln Glu Asp Glu Gly 575 580 585 Asn GlyAla Gln Tyr Arg Gly Ile Leu Glu Ala Ser Leu Glu Leu 590 595 600 Phe LysPhe Thr Ile Gly Met Gly Glu Leu Ala Phe Gln Glu Gln 605 610 615 Leu HisPhe Arg Gly Met Val Leu Leu Leu Leu Leu Ala Tyr Val 620 625 630 Leu LeuThr Tyr Ile Leu Leu Leu Asn Met Leu Ile Ala Leu Met 635 640 645 Ser GluThr Val Asn Ser Val Ala Thr Asp Ser Trp Ser Ile Trp 650 655 660 Lys LeuGln Lys Ala Ile Ser Val Leu Glu Met Glu Asn Gly Tyr 665 670 675 Trp TrpCys Arg Lys Lys Gln Arg Ala Gly Val Met Leu Thr Val 680 685 690 Gly ThrLys Pro Asp Gly Ser Pro Asp Glu Arg Trp Cys Phe Arg 695 700 705 Val GluGlu Val Asn Trp Ala Ser Trp Glu Gln Thr Leu Pro Thr 710 715 720 Leu CysGlu Asp Pro Ser Gly Ala Gly Val Pro Arg Thr Leu Glu 725 730 735 Asn ProVal Leu Ala Ser Pro Pro Lys Glu Asp Glu Asp Gly Ala 740 745 750 Ser GluGlu Asn Tyr Val Pro Val Gln Leu Leu Gln Ser Asn 755 760 4 255 PRT Homosapiens misc_feature Incyte ID No 2817822CD1 4 Met Trp Gln Gly Cys AlaVal Glu Arg Pro Val Gly Arg Met Thr 1 5 10 15 Ser Gln Thr Pro Leu ProGln Ser Pro Arg Pro Arg Arg Pro Thr 20 25 30 Met Ser Thr Val Val Glu LeuAsn Val Gly Gly Glu Phe His Thr 35 40 45 Thr Thr Leu Gly Thr Leu Arg LysPhe Pro Gly Ser Lys Leu Ala 50 55 60 Glu Met Phe Ser Ser Leu Ala Lys AlaSer Thr Asp Ala Glu Gly 65 70 75 Arg Phe Phe Ile Asp Arg Pro Ser Thr TyrPhe Arg Pro Ile Leu 80 85 90 Asp Tyr Leu Arg Thr Gly Gln Val Pro Thr GlnHis Ile Pro Glu 95 100 105 Val Tyr Arg Glu Ala Gln Phe Tyr Glu Ile LysPro Leu Val Lys 110 115 120 Leu Leu Glu Asp Met Pro Gln Ile Phe Gly GluGln Val Ser Arg 125 130 135 Lys Gln Phe Leu Leu Gln Val Pro Gly Tyr SerGlu Asn Leu Glu 140 145 150 Leu Met Val Arg Leu Ala Arg Ala Glu Ala IleThr Ala Arg Lys 155 160 165 Ser Ser Val Leu Val Cys Leu Val Glu Thr GluGlu Gln Asp Ala 170 175 180 Tyr Tyr Ser Glu Val Leu Cys Phe Leu Gln AspLys Lys Met Phe 185 190 195 Lys Ser Val Val Lys Phe Gly Pro Trp Lys AlaVal Leu Asp Asn 200 205 210 Ser Asp Leu Met His Cys Leu Glu Met Asp IleLys Ala Gln Gly 215 220 225 Tyr Lys Val Phe Ser Lys Phe Tyr Leu Thr TyrPro Thr Lys Arg 230 235 240 Asn Glu Phe His Phe Asn Ile Tyr Ser Phe ThrPhe Thr Trp Trp 245 250 255 5 584 PRT Homo sapiens misc_feature IncyteID No 4009329CD1 5 Met Ala Gly Arg Arg Leu Asn Leu Arg Trp Ala Leu SerVal Leu 1 5 10 15 Cys Val Leu Leu Met Ala Glu Thr Val Ser Gly Thr ArgGly Ser 20 25 30 Ser Thr Gly Ala His Ile Ser Pro Gln Phe Pro Ala Ser GlyVal 35 40 45 Asn Gln Thr Pro Val Val Asp Cys Arg Lys Val Cys Gly Leu Asn50 55 60 Val Ser Asp Arg Cys Asp Phe Ile Arg Thr Asn Pro Asp Cys His 6570 75 Ser Asp Gly Gly Tyr Leu Asp Tyr Leu Glu Gly Ile Phe Cys His 80 8590 Phe Pro Pro Ser Leu Leu Pro Leu Ala Val Thr Leu Tyr Val Ser 95 100105 Trp Leu Leu Tyr Leu Phe Leu Ile Leu Gly Val Thr Ala Ala Lys 110 115120 Phe Phe Cys Pro Asn Leu Ser Ala Ile Ser Thr Thr Leu Lys Leu 125 130135 Ser His Asn Val Ala Gly Val Thr Phe Leu Ala Phe Gly Asn Gly 140 145150 Ala Pro Asp Ile Phe Ser Ala Leu Val Ala Phe Ser Asp Pro His 155 160165 Thr Ala Gly Leu Ala Leu Gly Ala Leu Phe Gly Ala Gly Val Leu 170 175180 Val Thr Thr Val Val Ala Gly Gly Ile Thr Ile Leu His Pro Phe 185 190195 Met Ala Ala Ser Arg Pro Phe Phe Arg Asp Ile Val Phe Tyr Met 200 205210 Val Ala Val Phe Leu Thr Phe Leu Met Leu Phe Arg Gly Arg Val 215 220225 Thr Leu Ala Trp Ala Leu Gly Tyr Leu Gly Leu Tyr Val Phe Tyr 230 235240 Val Val Thr Val Ile Leu Cys Thr Trp Ile Tyr Gln Arg Gln Arg 245 250255 Arg Gly Ser Leu Phe Cys Pro Met Pro Val Thr Pro Glu Ile Leu 260 265270 Ser Asp Ser Glu Glu Asp Arg Val Ser Ser Asn Thr Asn Ser Tyr 275 280285 Asp Tyr Gly Asp Glu Tyr Arg Pro Leu Phe Phe Tyr Gln Glu Thr 290 295300 Thr Ala Gln Ile Leu Val Arg Ala Leu Asn Pro Leu Asp Tyr Met 305 310315 Lys Trp Arg Arg Lys Ser Ala Tyr Trp Lys Ala Leu Lys Val Phe 320 325330 Lys Leu Pro Val Glu Phe Leu Leu Leu Leu Thr Val Pro Val Val 335 340345 Asp Pro Asp Lys Asp Asp Gln Asn Trp Lys Arg Pro Leu Asn Cys 350 355360 Leu His Leu Val Ile Ser Pro Leu Val Val Val Leu Thr Leu Gln 365 370375 Ser Gly Thr Tyr Gly Val Tyr Glu Ile Gly Gly Leu Val Pro Val 380 385390 Trp Val Val Val Val Ile Ala Gly Thr Ala Leu Ala Ser Val Thr 395 400405 Phe Phe Ala Thr Ser Asp Ser Gln Pro Pro Arg Leu His Trp Leu 410 415420 Phe Ala Phe Leu Gly Phe Leu Thr Ser Ala Leu Trp Ile Asn Ala 425 430435 Ala Ala Thr Glu Val Val Asn Ile Leu Arg Ser Leu Gly Val Val 440 445450 Phe Arg Leu Ser Asn Thr Val Leu Gly Leu Thr Leu Leu Ala Trp 455 460465 Gly Asn Ser Ile Gly Asp Ala Phe Ser Asp Phe Thr Leu Ala Arg 470 475480 Gln Gly Tyr Pro Arg Met Ala Phe Ser Ala Cys Phe Gly Gly Ile 485 490495 Ile Phe Asn Ile Leu Val Gly Val Gly Leu Gly Cys Leu Leu Gln 500 505510 Ile Ser Arg Ser His Thr Glu Val Lys Leu Glu Pro Asp Gly Leu 515 520525 Leu Val Trp Val Leu Ala Gly Ala Leu Gly Leu Ser Leu Val Phe 530 535540 Ser Leu Val Ser Val Pro Leu Gln Cys Phe Gln Leu Ser Arg Val 545 550555 Tyr Gly Phe Cys Leu Leu Leu Phe Tyr Leu Asn Phe Leu Val Val 560 565570 Ala Leu Leu Ile Glu Phe Gly Val Ile His Leu Lys Ser Met 575 580 6416 PRT Homo sapiens misc_feature Incyte ID No 6618083CD1 6 Met Lys LeuSer Lys Lys Asp Arg Gly Glu Asp Glu Glu Ser Asp 1 5 10 15 Ser Ala LysLys Lys Leu Asp Trp Ser Cys Ser Leu Leu Val Ala 20 25 30 Ser Leu Ala GlyAla Phe Gly Ser Ser Phe Leu Tyr Gly Tyr Asn 35 40 45 Leu Ser Val Val AsnAla Pro Thr Pro Tyr Ile Lys Ala Phe Tyr 50 55 60 Asn Glu Ser Trp Glu ArgArg His Gly Arg Pro Ile Asp Pro Asp 65 70 75 Thr Leu Thr Leu Leu Trp SerVal Thr Val Ser Ile Phe Ala Ile 80 85 90 Gly Gly Leu Val Gly Thr Leu IleVal Lys Met Ile Gly Lys Val 95 100 105 Leu Gly Arg Lys His Thr Leu LeuAla Asn Asn Gly Phe Ala Ile 110 115 120 Ser Ala Ala Leu Leu Met Ala CysSer Leu Gln Ala Gly Ala Phe 125 130 135 Glu Met Leu Ile Val Gly Arg PheIle Met Gly Ile Asp Gly Gly 140 145 150 Val Ala Leu Ser Val Leu Pro MetTyr Leu Ser Glu Ile Ser Pro 155 160 165 Lys Glu Ile Arg Gly Ser Leu GlyGln Val Thr Ala Ile Phe Ile 170 175 180 Cys Ile Gly Val Phe Thr Gly GlnLeu Leu Gly Leu Pro Glu Leu 185 190 195 Leu Gly Lys Glu Ser Thr Trp ProTyr Leu Phe Gly Val Ile Val 200 205 210 Val Pro Ala Val Val Gln Leu LeuSer Leu Pro Phe Leu Pro Asp 215 220 225 Ser Pro Arg Tyr Leu Leu Leu GluLys His Asn Glu Ala Arg Ala 230 235 240 Val Lys Ala Phe Gln Thr Phe LeuGly Lys Ala Asp Val Ser Gln 245 250 255 Glu Val Glu Glu Val Leu Ala GluSer His Val Gln Arg Ser Ile 260 265 270 Arg Leu Val Ser Val Leu Glu LeuLeu Arg Ala Pro Tyr Val Arg 275 280 285 Trp Gln Val Val Thr Val Ile ValThr Met Ala Cys Tyr Gln Leu 290 295 300 Cys Gly Leu Asn Ala Ile Trp PheTyr Thr Asn Ser Ile Phe Gly 305 310 315 Lys Ala Gly Ile Pro Leu Ala LysIle Pro Tyr Val Thr Leu Ser 320 325 330 Thr Gly Gly Ile Glu Thr Leu AlaAla Val Phe Ser Gly Leu Val 335 340 345 Ile Glu His Leu Gly Arg Arg ProLeu Leu Ile Gly Gly Phe Gly 350 355 360 Leu Met Gly Leu Phe Phe Gly ThrLeu Thr Ile Thr Leu Thr Leu 365 370 375 Gln Asp His Ala Pro Trp Val ProTyr Leu Ser Ile Val Gly Ile 380 385 390 Leu Ala Ile Ile Ala Ser Phe CysSer Gly Pro Ala Val Phe Pro 395 400 405 Glu Glu Thr Val Asn Val Ser IleVal Ser Glu 410 415 7 664 PRT Homo sapiens misc_feature Incyte ID No7472002CD1 7 Met Thr Glu Lys Thr Asn Gly Val Lys Ser Ser Pro Ala Asn Asn1 5 10 15 His Asn His His Ala Pro Pro Ala Ile Lys Ala Asn Gly Lys Asp 2025 30 Asp His Arg Thr Ser Ser Arg Pro His Ser Ala Ala Asp Asp Asp 35 4045 Thr Ser Ser Glu Leu Gln Arg Leu Ala Asp Val Asp Ala Pro Gln 50 55 60Gln Gly Arg Ser Gly Phe Arg Arg Ile Val Arg Leu Val Gly Ile 65 70 75 IleArg Glu Trp Ala Asn Lys Asn Phe Arg Glu Glu Glu Pro Arg 80 85 90 Pro AspSer Phe Leu Glu Arg Phe Arg Gly Pro Glu Leu Gln Thr 95 100 105 Val ThrThr Gln Glu Gly Asp Gly Lys Gly Asp Lys Asp Gly Glu 110 115 120 Asp LysGly Thr Lys Lys Lys Phe Glu Leu Phe Val Leu Asp Pro 125 130 135 Ala GlyAsp Trp Tyr Tyr Cys Trp Leu Phe Val Ile Ala Met Pro 140 145 150 Val LeuTyr Asn Trp Cys Leu Leu Val Ala Arg Ala Cys Phe Ser 155 160 165 Asp LeuGln Lys Gly Tyr Tyr Leu Val Trp Leu Val Leu Asp Tyr 170 175 180 Val SerAsp Val Val Tyr Ile Ala Asp Leu Phe Ile Arg Leu Arg 185 190 195 Thr GlyPhe Leu Glu Gln Gly Leu Leu Val Lys Asp Thr Lys Lys 200 205 210 Leu ArgAsp Asn Tyr Ile His Thr Leu Gln Phe Lys Leu Asp Val 215 220 225 Ala SerIle Ile Pro Thr Asp Leu Ile Tyr Phe Ala Val Asp Ile 230 235 240 His SerPro Glu Val Arg Phe Asn Arg Leu Leu His Phe Ala Arg 245 250 255 Met PheGlu Phe Phe Asp Arg Thr Glu Thr Arg Thr Asn Tyr Pro 260 265 270 Asn IlePhe Arg Ile Ser Asn Leu Val Leu Tyr Ile Leu Val Ile 275 280 285 Ile HisTrp Asn Ala Cys Ile Tyr Tyr Ala Ile Ser Lys Ser Ile 290 295 300 Gly PheGly Val Asp Thr Trp Val Tyr Pro Asn Ile Thr Asp Pro 305 310 315 Glu TyrGly Tyr Leu Ala Arg Glu Tyr Ile Tyr Cys Leu Tyr Trp 320 325 330 Ser ThrLeu Thr Leu Thr Thr Ile Gly Glu Thr Pro Pro Pro Val 335 340 345 Lys AspGlu Glu Tyr Leu Phe Val Ile Phe Asp Phe Leu Ile Gly 350 355 360 Val LeuIle Phe Ala Thr Ile Val Gly Asn Val Gly Ser Met Ile 365 370 375 Ser AsnMet Asn Ala Thr Arg Ala Glu Phe Gln Ala Lys Ile Asp 380 385 390 Ala ValLys His Tyr Met Gln Phe Arg Lys Val Ser Lys Gly Met 395 400 405 Glu AlaLys Val Ile Arg Trp Phe Asp Tyr Leu Trp Thr Asn Lys 410 415 420 Lys ThrVal Asp Glu Arg Glu Ile Leu Lys Asn Leu Pro Ala Lys 425 430 435 Leu ArgAla Glu Ile Ala Ile Asn Val His Leu Ser Thr Leu Lys 440 445 450 Lys ValArg Ile Phe His Asp Cys Glu Ala Gly Leu Leu Val Glu 455 460 465 Leu ValLeu Lys Leu Arg Pro Gln Val Phe Ser Pro Gly Asp Tyr 470 475 480 Ile CysArg Lys Gly Asp Ile Gly Lys Glu Met Tyr Ile Ile Lys 485 490 495 Glu GlyLys Leu Ala Val Val Ala Asp Asp Gly Val Thr Gln Tyr 500 505 510 Ala LeuLeu Ser Ala Gly Ser Cys Phe Gly Glu Ile Ser Ile Leu 515 520 525 Asn IleLys Gly Ser Lys Met Gly Asn Arg Arg Thr Ala Asn Ile 530 535 540 Arg SerLeu Gly Tyr Ser Asp Leu Phe Cys Leu Ser Lys Asp Asp 545 550 555 Leu MetGlu Ala Val Thr Glu Tyr Pro Asp Ala Lys Lys Val Leu 560 565 570 Glu GluArg Gly Arg Glu Ile Leu Met Lys Glu Gly Leu Leu Asp 575 580 585 Glu AsnGlu Val Ala Thr Ser Met Glu Val Asp Val Gln Glu Lys 590 595 600 Leu GlyGln Leu Glu Thr Asn Met Glu Thr Leu Tyr Thr Arg Phe 605 610 615 Gly ArgLeu Leu Ala Glu Tyr Thr Gly Ala Gln Gln Lys Leu Lys 620 625 630 Gln ArgIle Thr Val Leu Glu Thr Lys Met Lys Gln Asn Asn Glu 635 640 645 Asp AspTyr Leu Ser Asp Gly Met Asn Ser Pro Glu Leu Ala Ala 650 655 660 Ala AspGlu Pro 8 242 PRT Homo sapiens misc_feature Incyte ID No 1812692CD1 8Met Ser Phe Arg Ala Ala Arg Leu Ser Met Arg Asn Arg Arg Asn 1 5 10 15Asp Thr Leu Asp Ser Thr Arg Thr Leu Tyr Ser Ser Ala Ser Arg 20 25 30 SerThr Asp Leu Ser Tyr Ser Glu Ser Asp Leu Val Asn Phe Ile 35 40 45 Gln AlaAsn Phe Lys Lys Arg Glu Cys Val Phe Phe Thr Lys Asp 50 55 60 Ser Lys AlaThr Glu Asn Val Cys Lys Cys Gly Tyr Ala Gln Ser 65 70 75 Gln His Met GluGly Thr Gln Ile Asn Gln Ser Glu Lys Trp Asn 80 85 90 Tyr Lys Lys His ThrLys Glu Phe Pro Thr Asp Ala Phe Gly Asp 95 100 105 Ile Gln Phe Glu ThrLeu Gly Lys Lys Gly Lys Tyr Ile Arg Leu 110 115 120 Ser Cys Asp Thr AspAla Glu Ile Leu Tyr Glu Leu Leu Thr Gln 125 130 135 His Trp His Leu LysThr Pro Asn Leu Val Ile Ser Val Thr Gly 140 145 150 Gly Ala Lys Asn PheAla Leu Lys Pro Arg Met Arg Lys Ile Phe 155 160 165 Ser Arg Leu Ile TyrIle Ala Gln Ser Lys Gly Ala Trp Ile Leu 170 175 180 Thr Gly Gly Thr HisTyr Gly Leu Met Lys Tyr Ile Gly Glu Val 185 190 195 Val Arg Asp Asn ThrIle Ser Arg Ser Ser Glu Glu Asn Ile Val 200 205 210 Ala Ile Gly Ile AlaAla Trp Gly Met Val Ser Asn Arg Asp Thr 215 220 225 Leu Ile Arg Asn CysAsp Ala Glu Val Pro Val Gly Gln Glu Glu 230 235 240 Val Cys 9 398 PRTHomo sapiens misc_feature Incyte ID No 3232992CD1 9 Met Val Ala Ala ProIle Phe Gly Tyr Leu Gly Asp Arg Phe Asn 1 5 10 15 Arg Lys Val Ile LeuSer Cys Gly Ile Phe Phe Trp Ser Ala Val 20 25 30 Thr Phe Ser Ser Ser PheIle Pro Gln Gln Tyr Phe Trp Leu Leu 35 40 45 Val Leu Ser Arg Gly Leu ValGly Ile Gly Glu Ala Ser Tyr Ser 50 55 60 Thr Ile Ala Pro Thr Ile Ile GlyAsp Leu Phe Thr Lys Asn Thr 65 70 75 Arg Thr Leu Met Leu Ser Val Phe TyrPhe Ala Ile Pro Leu Gly 80 85 90 Ser Gly Leu Gly Tyr Ile Thr Gly Ser SerVal Lys Gln Ala Ala 95 100 105 Gly Asp Trp His Trp Ala Leu Arg Val SerPro Val Leu Gly Met 110 115 120 Ile Thr Gly Thr Leu Ile Leu Ile Leu ValPro Ala Thr Lys Arg 125 130 135 Gly His Ala Asp Gln Leu Gly Asp Gln LeuLys Ala Arg Thr Ser 140 145 150 Trp Leu Arg Asp Met Lys Ala Leu Ile ArgAsn Arg Ser Tyr Val 155 160 165 Phe Ser Ser Leu Ala Thr Ser Ala Val SerPhe Ala Thr Gly Ala 170 175 180 Leu Gly Met Trp Ile Pro Leu Tyr Leu HisArg Ala Gln Val Val 185 190 195 Gln Lys Thr Ala Glu Thr Cys Asn Ser ProPro Cys Gly Ala Lys 200 205 210 Asp Ser Leu Ile Phe Gly Ala Ile Thr CysPhe Thr Gly Phe Leu 215 220 225 Gly Val Val Thr Gly Ala Gly Ala Thr ArgTrp Cys Arg Leu Lys 230 235 240 Thr Gln Arg Ala Asp Pro Leu Val Cys AlaVal Gly Met Leu Gly 245 250 255 Ser Ala Ile Phe Ile Cys Leu Ile Phe ValAla Ala Lys Ser Ser 260 265 270 Ile Val Gly Ala Tyr Ile Cys Ile Phe ValGly Glu Thr Leu Leu 275 280 285 Phe Ser Asn Trp Ala Ile Thr Ala Asp IleLeu Met Tyr Val Val 290 295 300 Ile Pro Thr Arg Arg Ala Thr Ala Val AlaLeu Gln Ser Phe Thr 305 310 315 Ser His Leu Leu Gly Asp Ala Gly Ser ProTyr Leu Ile Gly Phe 320 325 330 Ile Ser Asp Leu Ile Arg Gln Ser Thr LysAsp Ser Pro Leu Trp 335 340 345 Glu Phe Leu Ser Leu Gly Tyr Ala Leu MetLeu Cys Pro Phe Val 350 355 360 Val Val Leu Gly Gly Met Phe Phe Leu AlaThr Ala Leu Phe Phe 365 370 375 Val Ser Asp Arg Ala Arg Ala Glu Gln GlnVal Asn Gln Leu Ala 380 385 390 Met Pro Pro Ala Ser Val Lys Val 395 10553 PRT Homo sapiens misc_feature Incyte ID No 3358383CD1 10 Met Ala PheGln Asp Leu Leu Gly His Ala Gly Asp Leu Trp Arg 1 5 10 15 Phe Gln IleLeu Gln Thr Val Phe Leu Ser Ile Phe Ala Val Ala 20 25 30 Thr Tyr Leu HisPhe Met Leu Glu Asn Phe Thr Ala Phe Ile Pro 35 40 45 Gly His Arg Cys TrpVal His Ile Leu Asp Asn Asp Thr Val Ser 50 55 60 Asp Asn Asp Thr Gly AlaLeu Ser Gln Asp Ala Leu Leu Arg Ile 65 70 75 Ser Ile Pro Leu Asp Ser AsnMet Arg Pro Glu Lys Cys Arg Arg 80 85 90 Phe Val His Pro Gln Trp Gln LeuLeu His Leu Asn Gly Thr Phe 95 100 105 Pro Asn Thr Ser Asp Ala Asp MetGlu Pro Cys Val Asp Gly Trp 110 115 120 Val Tyr Asp Arg Ile Ser Phe SerSer Thr Ile Val Thr Glu Trp 125 130 135 Asp Leu Val Cys Asp Ser Gln SerLeu Thr Ser Val Ala Lys Phe 140 145 150 Val Phe Met Ala Gly Met Met ValGly Gly Ile Leu Gly Gly His 155 160 165 Leu Ser Asp Arg Phe Gly Arg ArgPhe Val Leu Arg Trp Cys Tyr 170 175 180 Leu Gln Val Ala Ile Val Gly ThrCys Ala Ala Leu Ala Pro Thr 185 190 195 Phe Leu Ile Tyr Cys Ser Leu ArgPhe Leu Ser Gly Ile Ala Ala 200 205 210 Met Ser Leu Ile Thr Asn Thr IleMet Leu Ile Ala Glu Trp Ala 215 220 225 Thr His Arg Phe Gln Ala Met GlyIle Thr Leu Gly Met Cys Pro 230 235 240 Ser Gly Ile Ala Phe Met Thr LeuAla Gly Leu Ala Phe Ala Ile 245 250 255 Arg Asp Trp His Ile Leu Gln LeuVal Val Ser Val Pro Tyr Phe 260 265 270 Val Ile Phe Leu Thr Ser Ser TrpLeu Leu Glu Ser Ala Arg Trp 275 280 285 Leu Ile Ile Asn Asn Lys Pro GluGlu Gly Leu Lys Glu Leu Arg 290 295 300 Lys Ala Ala His Arg Ser Gly MetLys Asn Ala Arg Asp Thr Leu 305 310 315 Thr Leu Glu Ile Leu Lys Ser ThrMet Lys Lys Glu Leu Glu Ala 320 325 330 Ala Gln Lys Lys Lys Pro Ser LeuCys Glu Met Leu His Met Pro 335 340 345 Asn Ile Cys Lys Arg Ile Ser LeuLeu Ser Phe Thr Arg Phe Ala 350 355 360 Asn Phe Met Ala Tyr Phe Gly LeuAsn Leu His Val Gln His Leu 365 370 375 Gly Asn Asn Val Phe Leu Leu GlnThr Leu Phe Gly Ala Val Ile 380 385 390 Leu Leu Ala Asn Cys Val Ala ProTrp Ala Leu Lys Tyr Met Thr 395 400 405 Arg Arg Ala Ser Gln Met Arg LeuMet Tyr Leu Leu Ala Ile Cys 410 415 420 Phe Met Ala Ile Ile Phe Val ProGln Glu Met Gln Thr Leu Arg 425 430 435 Glu Val Leu Ala Thr Leu Gly LeuGly Ala Ser Ala Leu Thr Asn 440 445 450 Thr Leu Ala Phe Ala His Gly AsnGlu Val Ile Pro Thr Ile Ile 455 460 465 Arg Ala Arg Ala Met Gly Ile AsnAla Thr Phe Ala Asn Ile Ala 470 475 480 Gly Ala Leu Ala Pro Leu Met MetIle Leu Ser Val Tyr Ser Pro 485 490 495 Pro Leu Pro Trp Ile Ile Tyr GlyVal Phe Pro Phe Ile Ser Gly 500 505 510 Phe Ala Phe Leu Leu Leu Pro GluThr Arg Asn Lys Pro Leu Phe 515 520 525 Asp Thr Ile Gln Asp Glu Lys AsnGlu Arg Lys Asp Pro Arg Glu 530 535 540 Pro Lys Gln Glu Asp Pro Arg ValGlu Val Thr Gln Phe 545 550 11 213 PRT Homo sapiens misc_feature IncyteID No 4250091CD1 11 Met Ser Ser Gln Glu Leu Val Thr Leu Asn Val Gly GlyLys Ile 1 5 10 15 Phe Thr Thr Arg Phe Ser Thr Ile Lys Gln Phe Pro AlaSer Arg 20 25 30 Leu Ala Arg Met Leu Asp Gly Arg Asp Gln Glu Phe Lys MetVal 35 40 45 Gly Gly Gln Ile Phe Val Asp Arg Asp Gly Asp Leu Phe Ser Phe50 55 60 Ile Leu Asp Phe Leu Arg Thr His Gln Leu Leu Leu Pro Thr Glu 6570 75 Phe Ser Asp Tyr Leu Arg Leu Gln Arg Glu Ala Leu Phe Tyr Glu 80 8590 Leu Arg Ser Leu Val Asp Leu Leu Asn Pro Tyr Leu Leu Gln Pro 95 100105 Arg Pro Ala Leu Val Glu Val His Phe Leu Ser Arg Asn Thr Gln 110 115120 Ala Phe Phe Arg Val Phe Gly Ser Cys Ser Lys Thr Ile Glu Met 125 130135 Leu Thr Gly Arg Ile Thr Val Phe Thr Glu Gln Pro Ser Ala Pro 140 145150 Thr Trp Asn Gly Asn Phe Phe Pro Pro Gln Met Thr Leu Leu Pro 155 160165 Leu Pro Pro Gln Arg Pro Ser Tyr His Asp Leu Val Phe Gln Cys 170 175180 Gly Ser Asp Ser Thr Thr Asp Asn Gln Thr Gly Val Arg Tyr Phe 185 190195 Val Leu Cys Ser Ile Ser Leu Val Tyr Gln Phe Val Met Phe Ser 200 205210 Leu Lys Thr 12 476 PRT Homo sapiens misc_feature Incyte ID No70064803CD1 12 Met Ala Gly Ser Asp Thr Ala Pro Phe Leu Ser Gln Ala AspAsp 1 5 10 15 Pro Asp Asp Gly Pro Val Pro Gly Thr Pro Gly Leu Pro GlySer 20 25 30 Thr Gly Asn Pro Lys Ser Glu Glu Pro Glu Val Pro Asp Gln Glu35 40 45 Gly Leu Gln Arg Ile Thr Gly Leu Ser Pro Gly Arg Ser Ala Leu 5055 60 Ile Val Ala Val Leu Cys Tyr Ile Asn Leu Leu Asn Tyr Met Asp 65 7075 Arg Phe Thr Val Ala Gly Val Leu Pro Asp Ile Glu Gln Phe Phe 80 85 90Asn Ile Gly Asp Ser Ser Ser Gly Leu Ile Gln Thr Val Phe Ile 95 100 105Ser Ser Tyr Met Val Leu Ala Pro Val Phe Gly Tyr Leu Gly Asp 110 115 120Arg Tyr Asn Arg Lys Tyr Leu Met Cys Gly Gly Ile Ala Phe Trp 125 130 135Ser Leu Val Thr Leu Gly Ser Ser Phe Ile Pro Gly Glu His Phe 140 145 150Trp Leu Leu Leu Leu Thr Arg Gly Leu Val Gly Val Gly Glu Ala 155 160 165Ser Tyr Ser Thr Ile Ala Pro Thr Leu Ile Ala Asp Leu Phe Val 170 175 180Ala Asp Gln Arg Ser Arg Met Leu Ser Ile Phe Tyr Phe Ala Ile 185 190 195Pro Val Gly Ser Gly Leu Gly Tyr Ile Ala Gly Ser Lys Val Lys 200 205 210Asp Met Ala Gly Asp Trp His Trp Ala Leu Arg Val Thr Pro Gly 215 220 225Leu Gly Val Val Ala Val Leu Leu Leu Phe Leu Val Val Arg Glu 230 235 240Pro Pro Arg Gly Ala Val Glu Arg His Ser Asp Leu Pro Pro Leu 245 250 255Asn Pro Thr Ser Trp Trp Ala Asp Leu Arg Ala Leu Ala Arg Asn 260 265 270Leu Ile Phe Gly Leu Ile Thr Cys Leu Thr Gly Val Leu Gly Val 275 280 285Gly Leu Gly Val Glu Ile Ser Arg Arg Leu Arg His Ser Asn Pro 290 295 300Arg Ala Asp Pro Leu Val Cys Ala Thr Gly Leu Leu Gly Ser Ala 305 310 315Pro Phe Leu Phe Leu Ser Leu Ala Cys Ala Arg Gly Ser Ile Val 320 325 330Ala Thr Tyr Ile Phe Ile Phe Ile Gly Glu Thr Leu Leu Ser Met 335 340 345Asn Trp Ala Ile Val Ala Asp Ile Leu Leu Tyr Val Val Ile Pro 350 355 360Thr Arg Arg Ser Thr Ala Glu Ala Phe Gln Ile Val Leu Ser His 365 370 375Leu Leu Gly Asp Ala Gly Ser Pro Tyr Leu Ile Gly Leu Ile Ser 380 385 390Asp Arg Leu Arg Arg Asn Trp Pro Pro Ser Phe Leu Ser Glu Phe 395 400 405Arg Ala Leu Gln Phe Ser Leu Met Leu Cys Ala Phe Val Gly Ala 410 415 420Leu Gly Gly Ala Ala Phe Leu Gly Thr Ala Ile Phe Ile Glu Ala 425 430 435Asp Arg Arg Arg Ala Gln Leu His Val Gln Gly Leu Leu His Glu 440 445 450Ala Gly Ser Thr Asp Asp Arg Ile Val Val Pro Gln Arg Gly Arg 455 460 465Ser Thr Arg Val Pro Val Ala Ser Val Leu Ile 470 475 13 246 PRT Homosapiens misc_feature Incyte ID No 70356768CD1 13 Met Leu His Ala Leu LeuArg Ser Arg Met Ile Gln Gly Arg Ile 1 5 10 15 Leu Leu Leu Thr Ile CysAla Ala Gly Ile Gly Gly Thr Phe Gln 20 25 30 Phe Gly Tyr Asn Leu Ser IleIle Asn Ala Pro Thr Leu His Ile 35 40 45 Gln Glu Phe Thr Asn Glu Thr TrpGln Ala Arg Thr Gly Glu Pro 50 55 60 Leu Pro Asp His Leu Val Leu Leu MetTrp Ser Leu Ile Val Ser 65 70 75 Leu Tyr Pro Leu Gly Gly Leu Phe Gly AlaLeu Leu Ala Gly Pro 80 85 90 Leu Ala Ile Thr Leu Gly Arg Lys Lys Ser LeuLeu Val Asn Asn 95 100 105 Ile Phe Val Val Ser Ala Ala Ile Leu Phe GlyPhe Ser Arg Lys 110 115 120 Ala Gly Ser Phe Glu Met Ile Met Leu Gly ArgLeu Leu Val Gly 125 130 135 Val Asn Ala Gly Val Ser Met Asn Ile Gln ProMet Tyr Leu Gly 140 145 150 Glu Ser Ala Pro Lys Glu Leu Arg Gly Ala ValAla Met Ser Ser 155 160 165 Ala Ile Phe Thr Ala Leu Gly Ile Val Met GlyGln Val Val Gly 170 175 180 Leu Arg Glu Leu Leu Gly Gly Pro Gln Ala TrpPro Leu Leu Leu 185 190 195 Ala Ser Cys Leu Val Pro Gly Ala Leu Gln LeuAla Ser Leu Pro 200 205 210 Leu Leu Pro Glu Ser Pro Arg Tyr Leu Leu IleAsp Cys Gly Asp 215 220 225 Thr Glu Ala Cys Leu Ala Glu Thr Gly Ser ArgLeu Ser Arg Leu 230 235 240 Glu Cys Cys Gly Cys Ser 245 14 436 PRT Homosapiens misc_feature Incyte ID No 5674114CD1 14 Met Gly Leu Ala Arg AlaLeu Arg Arg Leu Ser Gly Ala Leu Asp 1 5 10 15 Ser Gly Asp Ser Arg AlaGly Asp Glu Glu Glu Ala Gly Pro Gly 20 25 30 Leu Cys Arg Asn Gly Trp AlaPro Ala Pro Val Gln Ser Pro Val 35 40 45 Gly Arg Arg Arg Gly Arg Phe ValLys Lys Asp Gly His Cys Asn 50 55 60 Val Arg Phe Val Asn Leu Gly Gly GlnGly Ala Arg Tyr Leu Ser 65 70 75 Asp Leu Phe Thr Thr Cys Val Asp Val ArgTrp Arg Trp Met Cys 80 85 90 Leu Leu Phe Ser Cys Ser Phe Leu Ala Ser TrpLeu Leu Phe Gly 95 100 105 Leu Ala Phe Trp Leu Ile Ala Ser Leu His GlyAsp Leu Ala Ala 110 115 120 Pro Pro Pro Pro Ala Pro Cys Phe Ser His ValAla Ser Phe Leu 125 130 135 Ala Ala Phe Leu Phe Ala Leu Glu Thr Gln ThrSer Ile Gly Tyr 140 145 150 Gly Val Arg Ser Val Thr Glu Glu Cys Pro AlaAla Val Ala Ala 155 160 165 Val Val Leu Gln Cys Ile Ala Gly Cys Val LeuAsp Ala Phe Val 170 175 180 Val Gly Ala Val Met Ala Lys Met Ala Lys ProLys Lys Arg Asn 185 190 195 Glu Thr Leu Val Phe Ser Glu Asn Ala Val ValAla Leu Arg Asp 200 205 210 His Arg Leu Cys Leu Met Trp Arg Val Gly AsnLeu Arg Arg Ser 215 220 225 His Leu Val Glu Ala His Val Arg Ala Gln LeuLeu Gln Pro Arg 230 235 240 Val Thr Pro Glu Gly Glu Tyr Ile Pro Leu AspHis Gln Asp Val 245 250 255 Asp Val Gly Phe Asp Gly Gly Thr Asp Arg IlePhe Leu Val Ser 260 265 270 Pro Ile Thr Ile Val His Glu Ile Asp Ser AlaSer Pro Leu Tyr 275 280 285 Glu Leu Gly Arg Ala Glu Leu Ala Arg Ala AspPhe Glu Leu Val 290 295 300 Val Ile Leu Glu Gly Met Val Glu Ala Thr AlaMet Thr Thr Gln 305 310 315 Cys Arg Ser Ser Tyr Leu Pro Gly Glu Leu LeuTrp Gly His Arg 320 325 330 Phe Glu Pro Val Leu Phe Gln Arg Gly Ser GlnTyr Glu Val Asp 335 340 345 Tyr Arg His Phe His Arg Thr Tyr Glu Val ProGly Thr Pro Val 350 355 360 Cys Ser Ala Lys Glu Leu Asp Glu Arg Ala GluGln Ala Ser His 365 370 375 Ser Leu Lys Ser Ser Phe Pro Gly Ser Leu ThrAla Phe Cys Tyr 380 385 390 Glu Asn Glu Leu Ala Leu Ser Cys Cys Gln GluGlu Asp Glu Asp 395 400 405 Asp Glu Thr Glu Glu Gly Asn Gly Val Glu ThrGlu Asp Gly Ala 410 415 420 Ala Ser Pro Arg Val Leu Thr Pro Thr Leu AlaLeu Thr Leu Pro 425 430 435 Pro 15 453 PRT Homo sapiens misc_featureIncyte ID No 1254635CD1 15 Met Leu Lys Met Val Leu Thr Glu Asn Pro AsnGln Glu Ile Ala 1 5 10 15 Thr Ser Leu Glu Phe Leu Leu Leu Gln Asn SerPro Gly Ser Leu 20 25 30 Arg Ala Gln Gln Arg Met Ser Tyr Tyr Gly Ser SerTyr His Ile 35 40 45 Ile Asn Ala Asp Ala Lys Tyr Pro Gly Tyr Pro Pro GluHis Ile 50 55 60 Ile Ala Glu Lys Arg Arg Ala Arg Arg Arg Leu Leu His LysAsp 65 70 75 Gly Ser Cys Asn Val Tyr Phe Lys His Ile Phe Gly Glu Trp Gly80 85 90 Ser Tyr Val Val Asp Ile Phe Thr Thr Leu Val Asp Thr Lys Trp 95100 105 Arg His Met Phe Val Ile Phe Ser Leu Ser Tyr Ile Leu Ser Trp 110115 120 Leu Ile Phe Gly Ser Val Phe Trp Leu Ile Ala Phe His His Gly 125130 135 Asp Leu Leu Asn Asp Pro Asp Ile Thr Pro Cys Val Asp Asn Val 140145 150 His Ser Phe Thr Gly Ala Phe Leu Phe Ser Leu Glu Thr Gln Thr 155160 165 Thr Ile Gly Tyr Gly Tyr Arg Cys Val Thr Glu Glu Cys Ser Val 170175 180 Ala Val Leu Met Val Ile Leu Gln Ser Ile Leu Ser Cys Ile Ile 185190 195 Asn Thr Phe Ile Ile Gly Ala Ala Leu Ala Lys Met Ala Thr Ala 200205 210 Arg Lys Arg Ala Gln Thr Ile Arg Phe Ser Tyr Phe Ala Leu Ile 215220 225 Gly Met Arg Asp Gly Lys Leu Cys Leu Met Trp Arg Ile Gly Asp 230235 240 Phe Arg Pro Asn His Val Val Glu Gly Thr Val Arg Ala Gln Leu 245250 255 Leu Arg Tyr Thr Glu Asp Ser Glu Gly Arg Met Thr Met Ala Phe 260265 270 Lys Asp Leu Lys Leu Val Asn Asp Gln Ile Ile Leu Val Thr Pro 275280 285 Val Thr Ile Val His Glu Ile Asp His Glu Ser Pro Leu Tyr Ala 290295 300 Leu Asp Arg Lys Ala Val Ala Lys Asp Asn Phe Glu Ile Leu Val 305310 315 Thr Phe Ile Tyr Thr Gly Asp Ser Thr Gly Thr Ser His Gln Ser 320325 330 Arg Ser Ser Tyr Val Pro Arg Glu Ile Leu Trp Gly His Arg Phe 335340 345 Asn Asp Val Leu Glu Val Lys Arg Lys Tyr Tyr Lys Val Asn Cys 350355 360 Leu Gln Phe Glu Gly Ser Val Glu Val Tyr Ala Pro Phe Cys Ser 365370 375 Ala Lys Gln Leu Asp Trp Lys Asp Gln Gln Leu His Ile Glu Lys 380385 390 Ala Pro Pro Val Arg Glu Ser Cys Thr Ser Asp Thr Lys Ala Arg 395400 405 Arg Arg Ser Phe Ser Ala Val Ala Ile Val Ser Ser Cys Glu Asn 410415 420 Pro Glu Glu Thr Thr Thr Ser Ala Thr His Glu Tyr Arg Glu Thr 425430 435 Pro Tyr Gln Lys Ala Leu Leu Thr Leu Asn Arg Ile Ser Val Glu 440445 450 Ser Gln Met 16 299 PRT Homo sapiens misc_feature Incyte ID No1670595CD1 16 Met Ala Ser Glu Ser Ser Pro Leu Leu Ala Tyr Arg Leu LeuGly 1 5 10 15 Glu Glu Gly Val Ala Leu Pro Ala Asn Gly Ala Gly Gly ProGly 20 25 30 Gly Ala Ser Ala Arg Lys Leu Ser Thr Phe Leu Gly Val Val Val35 40 45 Pro Thr Val Leu Ser Met Phe Ser Ile Val Val Phe Leu Arg Ile 5055 60 Gly Phe Val Val Gly His Ala Gly Leu Leu Gln Ala Leu Ala Met 65 7075 Leu Leu Val Ala Tyr Phe Ile Leu Ala Leu Thr Val Leu Ser Val 80 85 90Cys Ala Ile Ala Thr Asn Gly Ala Val Gln Gly Gly Gly Ala Tyr 95 100 105Cys Ile Leu Gln His Arg Trp Thr Gly Met Pro Gln Gly Pro Val 110 115 120Gly Ser Gly Ser Cys Pro Arg Ala Thr Ala Trp Asn Leu Leu Tyr 125 130 135Gly Ser Leu Leu Leu Gly Leu Val Gly Gly Val Cys Thr Leu Gly 140 145 150Ala Gly Leu Tyr Ala Arg Ala Ser Phe Leu Thr Phe Leu Leu Val 155 160 165Ser Gly Ser Leu Ala Ser Val Leu Ile Ser Phe Val Ala Val Gly 170 175 180Pro Arg Asp Ile Arg Leu Thr Pro Arg Pro Gly Pro Asn Gly Ser 185 190 195Ser Leu Pro Pro Arg Phe Gly His Phe Thr Gly Phe Asn Ser Ser 200 205 210Thr Leu Lys Asp Asn Leu Gly Ala Gly Tyr Ala Glu Asp Tyr Thr 215 220 225Thr Gly Ala Val Met Asn Phe Ala Ser Val Phe Ala Val Leu Phe 230 235 240Asn Gly Arg His His Gly Trp Gly Gln His Val Arg Gly Ala Glu 245 250 255Gly Pro Gln Pro Gly Asp Pro Ser Gly His Asp Arg Arg Arg Arg 260 265 270Leu His Leu Leu Arg Leu Cys Pro Ala Phe Leu Ser Leu Gln Pro 275 280 285Pro Phe Thr Gly Ala Leu Met Leu Gly Ala Arg Pro Pro Leu 290 295 17 606PRT Homo sapiens misc_feature Incyte ID No 1859560CD1 17 Met Pro Ser SerVal Thr Ala Leu Gly Gln Ala Arg Ser Ser Gly 1 5 10 15 Pro Gly Met AlaPro Ser Ala Cys Cys Cys Ser Pro Ala Ala Leu 20 25 30 Gln Arg Arg Leu ProIle Leu Ala Trp Leu Pro Ser Tyr Ser Leu 35 40 45 Gln Trp Leu Lys Met AspPhe Val Ala Gly Leu Ser Val Gly Leu 50 55 60 Thr Ala Ile Pro Gln Ala LeuAla Tyr Ala Glu Val Ala Gly Leu 65 70 75 Pro Pro Gln Tyr Gly Leu Tyr SerAla Phe Met Gly Cys Phe Val 80 85 90 Tyr Phe Phe Leu Gly Thr Ser Arg AspVal Thr Leu Gly Pro Thr 95 100 105 Ala Ile Met Ser Leu Leu Val Ser PheTyr Thr Phe His Glu Pro 110 115 120 Ala Tyr Ala Val Leu Leu Ala Phe LeuSer Gly Cys Ile Gln Leu 125 130 135 Ala Met Gly Val Leu Arg Leu Gly PheLeu Leu Asp Phe Ile Ser 140 145 150 Tyr Pro Val Ile Lys Gly Phe Thr SerAla Ala Ala Val Thr Ile 155 160 165 Gly Phe Gly Gln Ile Lys Asn Leu LeuGly Leu Gln Asn Ile Pro 170 175 180 Arg Pro Phe Phe Leu Gln Val Tyr HisThr Phe Leu Arg Ile Ala 185 190 195 Glu Thr Arg Val Gly Asp Ala Val LeuGly Leu Val Cys Met Leu 200 205 210 Leu Leu Leu Val Leu Lys Leu Met ArgAsp His Val Pro Pro Val 215 220 225 His Pro Glu Met Pro Pro Gly Val ArgLeu Ser Arg Gly Leu Val 230 235 240 Trp Ala Ala Thr Thr Ala Arg Asn AlaLeu Val Val Ser Phe Ala 245 250 255 Ala Leu Val Ala Tyr Ser Phe Glu ValThr Gly Tyr Gln Pro Phe 260 265 270 Ile Leu Thr Gly Glu Thr Ala Glu GlyLeu Pro Pro Val Arg Ile 275 280 285 Pro Pro Phe Ser Val Thr Thr Ala AsnGly Thr Ile Ser Phe Thr 290 295 300 Glu Met Val Gln Asp Met Gly Ala GlyLeu Ala Val Val Pro Leu 305 310 315 Met Gly Leu Leu Glu Ser Ile Ala ValAla Lys Ala Phe Ala Ser 320 325 330 Gln Asn Asn Tyr Arg Ile Asp Ala AsnGln Glu Leu Leu Ala Ile 335 340 345 Gly Leu Thr Asn Met Leu Gly Ser LeuVal Ser Ser Tyr Pro Val 350 355 360 Thr Gly Ser Phe Gly Arg Thr Ala ValAsn Ala Gln Ser Gly Val 365 370 375 Cys Thr Pro Ala Gly Gly Leu Val ThrGly Val Leu Val Leu Leu 380 385 390 Ser Leu Asp Tyr Leu Thr Ser Leu PheTyr Tyr Ile Pro Lys Ser 395 400 405 Ala Leu Ala Ala Val Ile Ile Met AlaVal Ala Pro Leu Phe Asp 410 415 420 Thr Lys Ile Phe Arg Thr Leu Trp ArgVal Lys Arg Leu Asp Leu 425 430 435 Leu Pro Leu Cys Val Thr Phe Leu LeuCys Phe Trp Glu Val Gln 440 445 450 Tyr Gly Ile Leu Ala Gly Ala Leu ValSer Leu Leu Met Leu Leu 455 460 465 His Ser Ala Ala Arg Pro Glu Thr LysVal Ser Glu Gly Pro Val 470 475 480 Leu Val Leu Gln Pro Ala Ser Gly LeuSer Phe Pro Ala Met Glu 485 490 495 Ala Leu Arg Glu Glu Ile Leu Ser ArgAla Leu Glu Val Ser Pro 500 505 510 Pro Arg Cys Leu Val Leu Glu Cys ThrHis Val Cys Ser Ile Asp 515 520 525 Tyr Thr Val Val Leu Gly Leu Gly GluLeu Leu Gln Asp Phe Gln 530 535 540 Lys Gln Gly Val Ala Leu Ala Phe ValGly Leu Gln Val Pro Val 545 550 555 Leu Arg Val Leu Leu Ser Ala Asp LeuLys Gly Phe Gln Tyr Phe 560 565 570 Ser Thr Leu Glu Glu Ala Glu Lys HisLeu Arg Gln Glu Pro Gly 575 580 585 Thr Gln Pro Tyr Asn Ile Arg Glu AspSer Ile Leu Asp Gln Lys 590 595 600 Val Ala Leu Leu Lys Ala 605 18 324PRT Homo sapiens misc_feature Incyte ID No 5530164CD1 18 Met Ser Val GluAsp Gly Gly Met Pro Gly Leu Gly Arg Pro Arg 1 5 10 15 Gln Ala Arg TrpThr Leu Met Leu Leu Leu Ser Thr Ala Met Tyr 20 25 30 Gly Ala His Ala ProLeu Leu Ala Leu Cys His Val Asp Gly Arg 35 40 45 Val Pro Phe Arg Pro SerSer Ala Val Leu Leu Thr Glu Leu Thr 50 55 60 Lys Leu Leu Leu Cys Ala PheSer Leu Leu Val Gly Trp Gln Ala 65 70 75 Trp Pro Gln Gly Pro Pro Pro TrpArg Gln Ala Ala Pro Phe Ala 80 85 90 Leu Ser Ala Leu Leu Tyr Gly Ala AsnAsn Asn Leu Val Ile Tyr 95 100 105 Leu Gln Arg Tyr Met Asp Pro Ser ThrTyr Gln Val Leu Ser Asn 110 115 120 Leu Lys Ile Gly Ser Thr Ala Val LeuTyr Cys Leu Cys Leu Arg 125 130 135 His Arg Leu Ser Val Arg Gln Gly LeuAla Leu Leu Leu Leu Met 140 145 150 Ala Ala Gly Ala Cys Tyr Ala Ala GlyGly Leu Gln Val Pro Gly 155 160 165 Asn Thr Leu Pro Ser Pro Pro Pro AlaAla Ala Ala Ser Pro Met 170 175 180 Pro Leu His Ile Thr Pro Leu Gly LeuLeu Leu Leu Ile Leu Tyr 185 190 195 Cys Leu Ile Ser Gly Leu Ser Ser ValTyr Thr Glu Leu Leu Met 200 205 210 Lys Arg Gln Arg Leu Pro Leu Ala LeuGln Asn Leu Phe Leu Tyr 215 220 225 Thr Phe Gly Val Leu Leu Asn Leu GlyLeu His Ala Gly Gly Gly 230 235 240 Ser Gly Pro Gly Leu Leu Glu Gly PheSer Gly Trp Ala Ala Leu 245 250 255 Val Val Leu Ser Gln Ala Leu Asn GlyLeu Leu Met Ser Ala Val 260 265 270 Met Lys His Gly Ser Ser Ile Thr ArgLeu Phe Val Val Ser Cys 275 280 285 Ser Leu Val Val Asn Ala Val Leu SerAla Val Leu Leu Arg Leu 290 295 300 Gln Leu Thr Ala Ala Phe Phe Leu AlaThr Leu Leu Ile Gly Leu 305 310 315 Ala Met Arg Leu Tyr Tyr Gly Ser Arg320 19 445 PRT Homo sapiens misc_feature Incyte ID No 139115CD1 19 MetThr Leu Thr Gly Pro Leu Thr Thr Gln Tyr Val Tyr Arg Arg 1 5 10 15 IleTrp Glu Glu Thr Gly Asn Tyr Thr Phe Ser Ser Asp Ser Asn 20 25 30 Ile SerGlu Cys Glu Lys Asn Lys Ser Ser Pro Ile Phe Ala Phe 35 40 45 Gln Glu GluVal Gln Lys Lys Val Ser Arg Phe Asn Leu Gln Met 50 55 60 Asp Ile Ser GlyLeu Ile Pro Gly Leu Val Ser Thr Phe Ile Leu 65 70 75 Leu Ser Ile Ser AspHis Tyr Gly Arg Lys Phe Pro Met Ile Leu 80 85 90 Ser Ser Val Gly Ala LeuAla Thr Ser Val Trp Leu Cys Leu Leu 95 100 105 Cys Tyr Phe Ala Phe ProPhe Gln Leu Leu Ile Ala Ser Thr Phe 110 115 120 Ile Gly Ala Phe Cys GlyAsn Tyr Thr Thr Phe Trp Gly Ala Cys 125 130 135 Phe Ala Tyr Ile Val AspGln Cys Lys Glu His Lys Gln Lys Thr 140 145 150 Ile Arg Ile Ala Ile IleAsp Phe Leu Leu Gly Leu Val Thr Gly 155 160 165 Leu Thr Gly Leu Ser SerGly Tyr Phe Ile Arg Glu Leu Gly Phe 170 175 180 Glu Trp Ser Phe Leu IleIle Ala Val Ser Leu Ala Val Asn Leu 185 190 195 Ile Tyr Ile Leu Phe PheLeu Gly Asp Pro Val Lys Glu Cys Ser 200 205 210 Ser Gln Asn Val Thr MetSer Cys Ser Glu Gly Phe Lys Asn Leu 215 220 225 Phe Tyr Arg Thr Tyr MetLeu Phe Lys Asn Ala Ser Gly Lys Arg 230 235 240 Arg Phe Leu Leu Cys LeuLeu Leu Phe Thr Val Ile Thr Tyr Phe 245 250 255 Phe Val Val Ile Gly IleAla Pro Ile Phe Ile Leu Tyr Glu Leu 260 265 270 Asp Ser Pro Leu Cys TrpAsn Glu Val Phe Ile Gly Tyr Gly Ser 275 280 285 Ala Leu Gly Ser Ala SerPhe Leu Thr Ser Phe Leu Gly Ile Trp 290 295 300 Leu Phe Ser Tyr Cys MetGlu Asp Ile His Met Ala Phe Ile Gly 305 310 315 Ile Phe Thr Thr Met ThrGly Met Ala Met Thr Ala Phe Ala Ser 320 325 330 Thr Thr Leu Met Met PheLeu Ala Arg Val Pro Phe Leu Phe Thr 335 340 345 Ile Val Pro Phe Ser ValLeu Arg Ser Met Leu Ser Lys Val Val 350 355 360 Arg Ser Thr Glu Gln GlyThr Leu Phe Ala Cys Ile Ala Phe Leu 365 370 375 Glu Thr Leu Gly Gly ValThr Ala Val Ser Thr Phe Asn Gly Ile 380 385 390 Tyr Ser Ala Thr Val AlaTrp Tyr Pro Gly Phe Thr Phe Leu Leu 395 400 405 Ser Ala Gly Leu Leu LeuLeu Pro Ala Ile Ser Leu Cys Val Val 410 415 420 Lys Cys Thr Ser Trp AsnGlu Gly Ser Tyr Glu Leu Leu Ile Gln 425 430 435 Glu Glu Ser Ser Glu AspAla Ser Asp Arg 440 445 20 337 PRT Homo sapiens misc_feature Incyte IDNo 1702940CD1 20 Met Asn Pro Glu Ser Ser Ile Phe Ile Glu Asp Tyr Leu LysTyr 1 5 10 15 Phe Gln Asp Gln Val Ser Arg Glu Asn Leu Leu Gln Leu LeuThr 20 25 30 Asp Asp Glu Ala Trp Asn Gly Phe Val Ala Ala Ala Glu Leu Pro35 40 45 Arg Asp Glu Ala Asp Glu Leu Arg Lys Ala Leu Asn Lys Leu Ala 5055 60 Ser His Met Val Met Lys Asp Lys Asn Arg His Asp Lys Asp Gln 65 7075 Gln His Arg Gln Trp Phe Leu Lys Glu Phe Pro Arg Leu Lys Arg 80 85 90Glu Leu Glu Asp His Ile Arg Lys Leu Arg Ala Leu Ala Glu Glu 95 100 105Val Glu Gln Val His Arg Gly Thr Thr Ile Ala Asn Val Val Ser 110 115 120Asn Ser Val Gly Thr Thr Ser Gly Ile Leu Thr Leu Leu Gly Leu 125 130 135Gly Leu Ala Pro Phe Thr Glu Gly Ile Ser Phe Val Leu Leu Asp 140 145 150Thr Gly Met Gly Leu Gly Ala Ala Ala Ala Val Ala Gly Ile Thr 155 160 165Cys Ser Val Val Glu Leu Val Asn Lys Leu Arg Ala Arg Ala Gln 170 175 180Ala Arg Asn Leu Asp Gln Ser Gly Thr Asn Val Ala Lys Val Met 185 190 195Lys Glu Phe Val Gly Gly Asn Thr Pro Asn Val Leu Thr Leu Val 200 205 210Asp Asn Trp Tyr Gln Val Thr Gln Gly Ile Gly Arg Asn Ile Arg 215 220 225Ala Ile Arg Arg Ala Arg Ala Asn Pro Gln Leu Gly Ala Tyr Ala 230 235 240Pro Pro Pro His Val Ile Gly Arg Ile Ser Ala Glu Gly Gly Glu 245 250 255Gln Val Glu Arg Val Val Glu Gly Pro Ala Gln Ala Met Ser Arg 260 265 270Gly Thr Met Ile Val Gly Ala Ala Thr Gly Gly Ile Leu Leu Leu 275 280 285Leu Asp Val Val Ser Leu Ala Tyr Glu Ser Lys His Leu Leu Glu 290 295 300Gly Ala Lys Ser Glu Ser Ala Glu Glu Leu Lys Lys Arg Ala Gln 305 310 315Glu Leu Glu Gly Lys Leu Asn Phe Leu Thr Lys Ile His Glu Met 320 325 330Leu Gln Pro Gly Gln Asp Gln 335 21 273 PRT Homo sapiens misc_featureIncyte ID No 1703342CD1 21 Met Ala Thr Trp Asp Glu Lys Ala Val Thr ArgArg Ala Lys Val 1 5 10 15 Ala Pro Ala Glu Arg Met Ser Lys Phe Leu ArgHis Phe Thr Val 20 25 30 Val Gly Asp Asp Tyr His Ala Trp Asn Ile Asn TyrLys Lys Trp 35 40 45 Glu Asn Glu Glu Glu Glu Glu Glu Glu Glu Gln Pro ProPro Thr 50 55 60 Pro Val Ser Gly Glu Glu Gly Arg Ala Ala Ala Pro Asp ValAla 65 70 75 Pro Ala Pro Gly Pro Ala Pro Arg Ala Pro Leu Asp Phe Arg Gly80 85 90 Met Leu Arg Lys Leu Phe Ser Ser His Arg Phe Gln Val Ile Ile 95100 105 Ile Cys Leu Val Val Leu Asp Ala Leu Leu Val Leu Ala Glu Leu 110115 120 Ile Leu Asp Leu Lys Ile Ile Gln Pro Asp Lys Asn Asn Tyr Ala 125130 135 Ala Met Val Phe His Tyr Met Ser Ile Thr Ile Leu Val Phe Phe 140145 150 Met Met Glu Ile Ile Phe Lys Leu Phe Val Phe Arg Leu Glu Phe 155160 165 Phe His His Lys Phe Glu Ile Leu Asp Ala Val Val Val Val Val 170175 180 Ser Phe Ile Leu Asp Ile Val Leu Leu Phe Gln Glu His Gln Phe 185190 195 Glu Ala Leu Gly Leu Leu Ile Leu Leu Arg Leu Trp Arg Val Ala 200205 210 Arg Ile Ile Asn Gly Ile Ile Ile Ser Val Lys Thr Arg Ser Glu 215220 225 Arg Gln Leu Leu Arg Leu Lys Gln Met Asn Val Gln Leu Ala Ala 230235 240 Lys Ile Gln His Leu Glu Phe Ser Cys Ser Glu Lys Glu Gln Glu 245250 255 Ile Glu Arg Leu Asn Lys Leu Leu Arg Gln His Gly Leu Leu Gly 260265 270 Glu Val Asn 22 710 PRT Homo sapiens misc_feature Incyte ID No1727529CD1 22 Met Gly Gly Lys Gln Arg Asp Glu Asp Asp Glu Ala Tyr GlyLys 1 5 10 15 Pro Val Lys Tyr Asp Pro Ser Phe Arg Gly Pro Ile Lys AsnArg 20 25 30 Ser Cys Thr Asp Val Ile Cys Cys Val Leu Phe Leu Leu Phe Ile35 40 45 Leu Gly Tyr Ile Val Val Gly Ile Val Ala Trp Leu Tyr Gly Asp 5055 60 Pro Arg Gln Val Leu Tyr Pro Arg Asn Ser Thr Gly Ala Tyr Cys 65 7075 Gly Met Gly Glu Asn Lys Asp Lys Pro Tyr Leu Leu Tyr Phe Asn 80 85 90Ile Phe Ser Cys Ile Leu Ser Ser Asn Ile Ile Ser Val Ala Glu 95 100 105Asn Gly Leu Gln Cys Pro Thr Pro Gln Val Cys Val Ser Ser Cys 110 115 120Pro Glu Asp Pro Trp Thr Val Gly Lys Asn Glu Phe Ser Gln Thr 125 130 135Val Gly Glu Val Phe Tyr Thr Lys Asn Arg Asn Phe Cys Leu Pro 140 145 150Gly Val Pro Trp Asn Met Thr Val Ile Thr Ser Leu Gln Gln Glu 155 160 165Leu Cys Pro Ser Phe Leu Leu Pro Ser Ala Pro Ala Leu Gly Arg 170 175 180Cys Phe Pro Trp Thr Asn Ile Thr Pro Pro Ala Leu Pro Gly Ile 185 190 195Thr Asn Asp Thr Thr Ile Gln Gln Gly Ile Ser Gly Leu Ile Asp 200 205 210Ser Leu Asn Ala Arg Asp Ile Ser Val Lys Ile Phe Glu Asp Phe 215 220 225Ala Gln Ser Trp Tyr Trp Ile Leu Val Ala Leu Gly Val Ala Leu 230 235 240Val Leu Ser Leu Leu Phe Ile Leu Leu Leu Arg Leu Val Ala Gly 245 250 255Pro Leu Val Leu Val Leu Ile Leu Gly Val Leu Gly Val Leu Ala 260 265 270Tyr Gly Ile Tyr Tyr Cys Trp Glu Glu Tyr Arg Val Leu Arg Asp 275 280 285Lys Gly Ala Ser Ile Ser Gln Leu Gly Phe Thr Thr Asn Leu Ser 290 295 300Ala Tyr Gln Ser Val Gln Glu Thr Trp Leu Ala Ala Leu Ile Val 305 310 315Leu Ala Val Leu Glu Ala Ile Leu Leu Leu Val Leu Ile Phe Leu 320 325 330Arg Gln Arg Ile Arg Ile Ala Ile Ala Leu Leu Lys Glu Ala Ser 335 340 345Lys Ala Val Gly Gln Met Met Ser Thr Met Phe Tyr Pro Leu Val 350 355 360Thr Phe Val Leu Leu Leu Ile Cys Ile Ala Tyr Trp Ala Met Thr 365 370 375Ala Leu Tyr Leu Ala Thr Ser Gly Gln Pro Gln Tyr Val Leu Trp 380 385 390Ala Ser Asn Ile Ser Ser Pro Gly Cys Glu Lys Val Pro Ile Asn 395 400 405Thr Ser Cys Asn Pro Thr Ala His Leu Val Asn Ser Ser Cys Pro 410 415 420Gly Leu Met Cys Val Phe Gln Gly Tyr Ser Ser Lys Gly Leu Ile 425 430 435Gln Arg Ser Val Phe Asn Leu Gln Ile Tyr Gly Val Leu Gly Leu 440 445 450Phe Trp Thr Leu Asn Trp Val Leu Ala Leu Gly Gln Cys Val Leu 455 460 465Ala Gly Ala Phe Ala Ser Phe Tyr Trp Ala Phe His Lys Pro Gln 470 475 480Asp Ile Pro Thr Phe Pro Leu Ile Ser Ala Phe Ile Arg Thr Leu 485 490 495Arg Tyr His Thr Gly Ser Leu Ala Phe Gly Ala Leu Ile Leu Thr 500 505 510Leu Val Gln Ile Ala Arg Val Ile Leu Glu Tyr Ile Asp His Lys 515 520 525Leu Arg Gly Val Gln Asn Pro Val Ala Arg Cys Ile Met Cys Cys 530 535 540Phe Lys Cys Cys Leu Trp Cys Leu Glu Lys Phe Ile Lys Phe Leu 545 550 555Asn Arg Asn Ala Tyr Ile Met Ile Ala Ile Tyr Gly Lys Asn Phe 560 565 570Cys Val Ser Ala Lys Asn Ala Phe Met Leu Leu Met Arg Asn Ile 575 580 585Val Arg Val Val Val Leu Asp Lys Val Thr Asp Leu Leu Leu Phe 590 595 600Phe Gly Lys Leu Leu Val Val Gly Gly Val Gly Val Leu Ser Phe 605 610 615Phe Phe Phe Ser Gly Arg Ile Pro Gly Leu Gly Lys Asp Phe Lys 620 625 630Ser Pro His Leu Asn Tyr Tyr Trp Leu Pro Ile Met Thr Ser Ile 635 640 645Leu Gly Ala Tyr Val Ile Ala Ser Gly Phe Phe Ser Val Phe Gly 650 655 660Met Cys Val Asp Thr Leu Phe Leu Cys Phe Leu Glu Asp Leu Glu 665 670 675Arg Asn Asn Gly Ser Leu Asp Arg Pro Tyr Tyr Met Ser Lys Ser 680 685 690Leu Leu Lys Ile Leu Gly Lys Lys Asn Glu Ala Pro Pro Asp Asn 695 700 705Lys Lys Arg Lys Lys 710 23 476 PRT Homo sapiens misc_feature Incyte IDNo 2289333CD1 23 Glu Gln Asn Phe Asp Gly Thr Ser Asp Glu Glu His Glu GlnGlu 1 5 10 15 Leu Leu Pro Val Gln Lys His Tyr Gln Leu Asp Asp Gln GluGly 20 25 30 Ile Ser Phe Val Gln Thr Leu Met His Leu Leu Lys Gly Asn Ile35 40 45 Gly Thr Gly Leu Leu Gly Leu Pro Leu Ala Ile Lys Asn Ala Gly 5055 60 Ile Val Leu Gly Pro Ile Ser Leu Val Phe Ile Gly Ile Ile Ser 65 7075 Val His Cys Met His Ile Leu Val Arg Cys Ser His Phe Leu Cys 80 85 90Leu Arg Phe Lys Lys Ser Thr Leu Gly Tyr Ser Asp Thr Val Ser 95 100 105Phe Ala Met Glu Val Ser Pro Trp Ser Cys Leu Gln Lys Gln Ala 110 115 120Ala Trp Gly Arg Ser Val Val Asp Phe Phe Leu Val Ile Thr Gln 125 130 135Leu Gly Phe Cys Ser Val Tyr Ile Val Phe Leu Ala Glu Asn Val 140 145 150Lys Gln Val His Glu Gly Phe Leu Glu Ser Lys Val Phe Ile Ser 155 160 165Asn Ser Thr Asn Ser Ser Asn Pro Cys Glu Arg Arg Ser Val Asp 170 175 180Leu Arg Ile Tyr Met Leu Cys Phe Leu Pro Phe Ile Ile Leu Leu 185 190 195Val Phe Ile Arg Glu Leu Lys Asn Leu Phe Val Leu Ser Phe Leu 200 205 210Ala Asn Val Ser Met Ala Val Ser Leu Val Ile Ile Tyr Gln Tyr 215 220 225Val Val Arg Asn Met Pro Asp Pro His Asn Leu Pro Ile Val Ala 230 235 240Gly Trp Lys Lys Tyr Pro Leu Phe Phe Gly Thr Ala Val Phe Ala 245 250 255Phe Glu Gly Ile Gly Val Val Leu Pro Leu Glu Asn Gln Met Lys 260 265 270Glu Ser Lys Arg Phe Pro Gln Ala Leu Asn Ile Gly Met Gly Ile 275 280 285Val Thr Thr Leu Tyr Val Thr Leu Ala Thr Leu Gly Tyr Met Cys 290 295 300Phe His Asp Glu Ile Lys Gly Ser Ile Thr Leu Asn Leu Pro Gln 305 310 315Asp Val Trp Leu Tyr Gln Ser Val Lys Ile Leu Tyr Ser Phe Gly 320 325 330Ile Phe Val Thr Tyr Ser Ile Gln Phe Tyr Val Pro Ala Glu Ile 335 340 345Ile Ile Pro Gly Ile Thr Ser Lys Phe His Thr Lys Trp Lys Gln 350 355 360Ile Cys Glu Phe Gly Ile Arg Ser Phe Leu Val Ser Ile Thr Cys 365 370 375Ala Gly Ala Ile Leu Ile Pro Arg Leu Asp Ile Val Ile Ser Phe 380 385 390Val Gly Ala Val Ser Ser Ser Thr Leu Ala Leu Ile Leu Pro Pro 395 400 405Leu Val Glu Ile Leu Thr Phe Ser Lys Glu His Tyr Asn Ile Trp 410 415 420Met Val Leu Lys Asn Ile Ser Ile Ala Phe Thr Gly Val Val Gly 425 430 435Phe Leu Leu Gly Thr Tyr Ile Thr Val Glu Glu Ile Ile Tyr Pro 440 445 450Thr Pro Lys Val Val Ala Gly Thr Pro Gln Ser Pro Phe Leu Asn 455 460 465Leu Asn Ser Thr Cys Leu Thr Ser Gly Leu Lys 470 475 24 237 PRT Homosapiens misc_feature Incyte ID No 2720354CD1 24 Met Gly Leu Thr Phe IleAsn Ala Leu Val Phe Gly Val Gln Gly 1 5 10 15 Asn Thr Leu Arg Ala LeuGly His Asp Ser Pro Leu Asn Gln Phe 20 25 30 Leu Ala Gly Ala Ala Ala GlyAla Ile Gln Cys Val Ile Cys Cys 35 40 45 Pro Met Glu Leu Ala Lys Thr ArgLeu Gln Leu Gln Asp Ala Gly 50 55 60 Pro Ala Arg Thr Tyr Lys Gly Ser LeuAsp Cys Leu Ala Gln Ile 65 70 75 Tyr Gly His Glu Gly Leu Arg Gly Val AsnArg Gly Met Val Ser 80 85 90 Thr Leu Leu Arg Glu Thr Pro Ser Phe Gly ValTyr Phe Leu Thr 95 100 105 Tyr Asp Ala Leu Thr Arg Ala Leu Gly Cys GluPro Gly Asp Arg 110 115 120 Leu Leu Val Pro Lys Leu Leu Leu Ala Gly GlyThr Ser Gly Ile 125 130 135 Val Ser Trp Leu Ser Thr Tyr Pro Val Asp ValVal Lys Ser Arg 140 145 150 Leu Gln Ala Asp Gly Leu Arg Gly Ala Pro ArgTyr Arg Gly Ile 155 160 165 Leu Asp Cys Val His Gln Ser Tyr Arg Ala GluGly Trp Arg Val 170 175 180 Phe Thr Arg Gly Leu Ala Ser Thr Leu Leu ArgAla Phe Pro Val 185 190 195 Asn Ala Ala Thr Phe Ala Thr Val Thr Val ValLeu Thr Tyr Ala 200 205 210 Arg Gly Glu Glu Ala Gly Pro Glu Gly Glu AlaVal Pro Ala Ala 215 220 225 Pro Ala Gly Pro Ala Leu Ala Gln Pro Ser SerLeu 230 235 25 345 PRT Homo sapiens misc_feature Incyte ID No 3038193CD125 Met Arg Leu Leu Glu Arg Met Arg Lys Asp Trp Phe Met Val Gly 1 5 10 15Ile Val Leu Ala Ile Ala Gly Ala Lys Leu Glu Pro Ser Ile Gly 20 25 30 ValAsn Gly Gly Pro Leu Lys Pro Glu Ile Thr Val Ser Tyr Ile 35 40 45 Ala ValAla Thr Ile Phe Phe Asn Ser Gly Leu Ser Leu Lys Thr 50 55 60 Glu Glu LeuThr Ser Ala Leu Val His Leu Lys Leu His Leu Phe 65 70 75 Ile Gln Ile PheThr Leu Ala Phe Phe Pro Ala Thr Ile Trp Leu 80 85 90 Phe Leu Gln Leu LeuSer Ile Thr Pro Ile Asn Glu Trp Leu Leu 95 100 105 Lys Gly Leu Gln ThrVal Gly Cys Met Pro Pro Pro Val Ser Ser 110 115 120 Ala Val Ile Leu ThrLys Ala Val Gly Gly Asn Glu Gly Ile Val 125 130 135 Ile Thr Pro Leu LeuLeu Leu Leu Phe Leu Gly Ser Ser Ser Ser 140 145 150 Val Pro Phe Thr SerIle Phe Ser Gln Leu Phe Met Thr Val Val 155 160 165 Val Pro Leu Ile IleGly Gln Ile Val Arg Arg Tyr Ile Lys Asp 170 175 180 Trp Leu Glu Arg LysLys Pro Pro Phe Gly Ala Ile Ser Ser Ser 185 190 195 Val Leu Leu Met IleIle Tyr Thr Thr Phe Cys Asp Thr Phe Ser 200 205 210 Asn Pro Asn Ile AspLeu Asp Lys Phe Ser Leu Val Leu Ile Leu 215 220 225 Phe Ile Ile Phe SerIle Gln Leu Ser Phe Met Leu Leu Thr Phe 230 235 240 Ile Phe Ser Thr ArgAsn Asn Ser Gly Phe Thr Pro Ala Asp Thr 245 250 255 Val Ala Ile Ile PheCys Ser Thr His Lys Ser Leu Thr Leu Gly 260 265 270 Ile Pro Met Leu LysIle Val Phe Ala Gly Tyr Glu His Leu Ser 275 280 285 Leu Ile Ser Val ProLeu Leu Ile Tyr His Pro Ala Gln Ile Leu 290 295 300 Leu Gly Ser Val LeuVal Pro Thr Ile Lys Ser Trp Met Val Ser 305 310 315 Arg Gln Lys Lys LeuLeu Gln Thr Arg Gly Pro Leu Ala Asn Leu 320 325 330 Asn Asn Pro Glu GlyLeu Glu Tyr Leu Ser Ile Lys Phe Gly His 335 340 345 26 521 PRT Homosapiens misc_feature Incyte ID No 3460979CD1 26 Met Ala Ala Leu Ala ProVal Gly Ser Pro Ala Ser Arg Gly Pro 1 5 10 15 Arg Leu Ala Ala Gly LeuArg Leu Leu Pro Met Leu Gly Leu Leu 20 25 30 Gln Leu Leu Ala Glu Pro GlyLeu Gly Arg Val His His Leu Ala 35 40 45 Leu Lys Asp Asp Val Arg His LysVal His Leu Asn Thr Phe Gly 50 55 60 Phe Phe Lys Asp Gly Tyr Met Val ValAsn Val Ser Ser Leu Ser 65 70 75 Leu Asn Glu Pro Glu Asp Lys Asp Val ThrIle Gly Phe Ser Leu 80 85 90 Asp Arg Thr Lys Asn Asp Gly Phe Ser Ser TyrLeu Asp Glu Asp 95 100 105 Val Asn Tyr Cys Ile Leu Lys Lys Gln Ser ValSer Val Thr Leu 110 115 120 Leu Ile Leu Asp Ile Ser Arg Ser Glu Val ArgVal Lys Ser Pro 125 130 135 Pro Glu Ala Gly Thr Gln Leu Pro Lys Ile IlePhe Ser Arg Asp 140 145 150 Glu Lys Val Leu Gly Gln Ser Gln Glu Pro AsnVal Asn Pro Ala 155 160 165 Ser Ala Gly Asn Gln Thr Gln Lys Thr Gln AspGly Gly Lys Ser 170 175 180 Lys Arg Ser Thr Val Asp Ser Lys Ala Met GlyGlu Lys Ser Phe 185 190 195 Ser Val His Asn Asn Gly Gly Ala Val Ser PheGln Phe Phe Phe 200 205 210 Asn Ile Ser Thr Asp Asp Gln Glu Gly Leu TyrSer Leu Tyr Phe 215 220 225 His Lys Cys Leu Gly Lys Glu Leu Pro Ser AspLys Phe Thr Phe 230 235 240 Ser Leu Asp Ile Glu Ile Thr Glu Lys Asn ProAsp Ser Tyr Leu 245 250 255 Ser Ala Gly Glu Ile Pro Leu Pro Lys Leu TyrIle Ser Met Ala 260 265 270 Phe Phe Phe Phe Leu Ser Gly Thr Ile Trp IleHis Ile Leu Arg 275 280 285 Lys Arg Arg Asn Asp Val Phe Lys Ile His TrpLeu Met Ala Ala 290 295 300 Leu Pro Phe Thr Lys Ser Leu Ser Leu Val PheHis Ala Ile Asp 305 310 315 Tyr His Tyr Ile Ser Ser Gln Gly Phe Pro IleGlu Gly Trp Ala 320 325 330 Val Val Tyr Tyr Ile Thr His Leu Leu Lys GlyAla Leu Leu Phe 335 340 345 Ile Thr Ile Ala Leu Ile Gly Thr Gly Trp AlaPhe Ile Lys His 350 355 360 Ile Leu Ser Asp Lys Asp Lys Lys Ile Phe MetIle Val Ile Pro 365 370 375 Leu Gln Val Leu Ala Asn Val Ala Tyr Ile IleIle Glu Ser Thr 380 385 390 Glu Glu Gly Thr Thr Glu Tyr Gly Leu Trp LysAsp Ser Leu Phe 395 400 405 Leu Val Asp Leu Leu Cys Cys Gly Ala Ile LeuPhe Pro Val Val 410 415 420 Trp Ser Ile Arg His Leu Gln Glu Ala Ser AlaThr Asp Gly Lys 425 430 435 Ala Ala Ile Asn Leu Ala Lys Leu Lys Leu PheArg His Tyr Tyr 440 445 450 Val Leu Ile Val Cys Tyr Ile Tyr Phe Thr ArgIle Ile Ala Phe 455 460 465 Leu Leu Lys Leu Ala Val Pro Phe Gln Trp LysTrp Leu Tyr Gln 470 475 480 Leu Leu Asp Glu Thr Ala Thr Leu Val Phe PheVal Leu Thr Gly 485 490 495 Tyr Lys Phe Arg Pro Ala Ser Asp Asn Pro TyrLeu Gln Leu Ser 500 505 510 Gln Glu Glu Glu Asp Leu Glu Met Glu Ser Val515 520 27 555 PRT Homo sapiens misc_feature Incyte ID No 7472200CD1 27Met Thr Leu Val Tyr Phe Pro Pro Ser Lys Leu Gln Gln Gln Gln 1 5 10 15Gln Pro Ser Arg Ser Ser Arg Leu Ala Gln Gln Leu Ala Gln Ser 20 25 30 SerTrp Gln Leu Ala Leu Arg Phe Gly Lys Arg Thr Thr Ile His 35 40 45 Gly LeuAsp Arg Leu Leu Ser Ala Lys Ala Ser Arg Trp Glu Arg 50 55 60 Phe Val TrpLeu Cys Thr Phe Val Ser Ala Phe Leu Gly Ala Val 65 70 75 Tyr Val Cys LeuIle Leu Ser Ala Arg Tyr Asn Ala Ala His Phe 80 85 90 Gln Thr Val Val AspSer Thr Arg Phe Pro Val Tyr Arg Ile Pro 95 100 105 Phe Pro Val Ile ThrIle Cys Asn Arg Asn Arg Leu Asn Trp Gln 110 115 120 Arg Leu Ala Glu AlaLys Ser Arg Phe Leu Ala Asn Gly Ser Asn 125 130 135 Ser Ala Gln Gln GluLeu Phe Glu Leu Ile Val Gly Thr Tyr Asp 140 145 150 Asp Ala Tyr Phe GlyHis Phe Gln Ser Phe Glu Arg Leu Arg Asn 155 160 165 Gln Pro Thr Glu LeuLeu Asn Tyr Val Asn Phe Ser Gln Val Val 170 175 180 Asp Phe Met Thr TrpArg Cys Asn Glu Leu Leu Ala Glu Cys Leu 185 190 195 Trp Arg His His AlaTyr Asp Cys Cys Glu Ile Arg Ser Lys Arg 200 205 210 Arg Ser Lys Asn GlyLeu Cys Trp Ala Phe Asn Ser Leu Glu Thr 215 220 225 Glu Glu Gly Arg ArgMet Gln Leu Leu Asp Pro Met Trp Pro Trp 230 235 240 Arg Thr Gly Ser AlaGly Pro Met Ser Ala Leu Ser Val Arg Val 245 250 255 Leu Ile Gln Pro AlaLys His Trp Pro Gly His Arg Glu Thr Asn 260 265 270 Ala Met Lys Gly IleAsp Val Met Val Thr Glu Pro Phe Val Trp 275 280 285 His Asn Asn Pro PhePhe Val Ala Ala Asn Thr Glu Thr Thr Met 290 295 300 Glu Ile Glu Pro ValIle Tyr Phe Tyr Asp Asn Asp Thr Arg Gly 305 310 315 Val Arg Ser Asp GlnArg Gln Cys Val Phe Asp Asp Glu His Asn 320 325 330 Ser Lys Asp Phe LysSer Leu Gln Gly Tyr Val Tyr Met Ile Glu 335 340 345 Asn Cys Gln Ser GluCys His Gln Glu Tyr Leu Val Arg Tyr Cys 350 355 360 Asn Cys Thr Met AspLeu Leu Phe Pro Pro Asp Leu Leu Ile Tyr 365 370 375 Ser His Asn Pro GlyGlu Lys Glu Phe Val Arg Asn Gln Phe Gln 380 385 390 Gly Met Ser Cys LysCys Phe Arg Asn Cys Tyr Ser Leu Asn Tyr 395 400 405 Ile Ser Asp Val ArgPro Ala Phe Leu Pro Pro Asp Val Tyr Ala 410 415 420 Asn Asn Ser Tyr ValAsp Leu Asp Val His Phe Arg Phe Glu Thr 425 430 435 Ile Met Val Tyr ArgThr Ser Leu Val Phe Gly Trp Val Asp Leu 440 445 450 Met Val Ser Phe GlyGly Ile Ala Gly Leu Phe Leu Gly Cys Ser 455 460 465 Leu Ile Ser Gly MetGlu Leu Ala Tyr Phe Leu Cys Ile Glu Val 470 475 480 Pro Ala Phe Gly LeuAsp Gly Leu Arg Arg Arg Trp Lys Ala Arg 485 490 495 Arg Gln Met Asp LeuGly Val Thr Val Pro Thr Pro Thr Leu Asn 500 505 510 Phe Gln Gln Thr ThrPro Ser Gln Leu Met Glu Asn Tyr Ile Met 515 520 525 Gln Leu Lys Ala GluLys Ala Gln Gln Gln Lys Ala Asn Phe Gln 530 535 540 Asn Trp His Arg IleThr Phe Ala Gln Lys His Val Ile Gly Lys 545 550 555 28 2080 DNA Homosapiens misc_feature Incyte ID No 1416107CB1 28 ggcggttcag gcgccagagctggccgatcg gcgttggccg ccgacatgac gcccgaggac 60 ccagaggaaa cccagccgcttctggggcct cctggcggca gcgcgccccg cggccgccgc 120 gtcttcctcg ccgccttcgccgctgccctg ggcccactca gcttcggctt cgcgctcggc 180 tacagctccc cggccatccctagcctgcag cgcgccgcgc ccccggcccc gcgcctggac 240 gacgccgccg cctcctggttcggggctgtc gtgaccctgg gtgccgcggc ggggggagtg 300 ctgggcggct ggctggtggaccgcgccggg cgcaagctga gcctcttgct gtgctccgtg 360 cccttcgtgg ccggctttgccgtcatcacc gcggcccagg acgtgtggat gctgctgggg 420 ggccgcctcc tcaccggcctggcctgcggt gttgcctccc tagtggcccc ggtctacatc 480 tccgaaatcg cctacccagcagtccggggg ttgctcggct cctgtgtgca gctaatggtc 540 gtcgtcggca tcctcctggcctacctggca ggctgggtgc tggagtggcg ctggctggct 600 gtgctgggct gcgtgcccccctccctcatg ctgcttctca tgtgcttcat gcccgagacc 660 ccgcgcttcc tgctgactcagcacaggcgc caggaggcca tggccgccct gcggttcctg 720 tggggctccg agcagggctgggaagacccc cccatcgggg ctgagcagag ctttcacctg 780 gccctgctgc ggcagcccggcatctacaag cccttcatca tcggcgtctc cctgatggcc 840 ttccagcagc tgtcgggggtcaacgccgtc atgttctatg cagagaccat ctttgaagag 900 gccaagttca aggacagcagcctggcctcg gtcgtcgtgg gtgtcatcca ggtgctgttc 960 acagctgtgg cggctctcatcatggacaga gcagggcgga ggctgctcct ggtcttgtca 1020 ggtgtggtca tggtgttcagcacgagtgcc ttcggcgcct acttcaagct gacccagggt 1080 ggccctggca actcctcgcacgtggccatc tcggcgcctg tctctgcaca gcctgttgat 1140 gccagcgtgg ggctggcctggctggccgtg ggcagcatgt gcctcttcat cgccggcttt 1200 gcggtgggct gggggcccatcccctggctc ctcatgtcag agatcttccc tctgcatgtc 1260 aagggcgtgg cgacaggcatctgcgtcctc accaactggc tcatggcctt tctcgtgacc 1320 aaggagttca gcagcctcatggaggtcctc aggccctatg gagccttctg gcttgcctcc 1380 gctttctgca tcttcagtgtccttttcact ttgttctgtg tccctgaaac taaaggaaag 1440 actctggaac aaatcacagcccattttgag gggcgatgac agccactcac taggggatgg 1500 agcaagcctg tgactccaagctgggcccaa gcccagagcc cctgcctgcc ccaggggagc 1560 cagaatccag ccccttggagccttggtctg cagggtccct ccttcctgtc atgctccctc 1620 cagcccatga cccggggctaggaggctcac tgcctcctgt tccagctcct gctgctgctc 1680 tgaggactca ggaacaccttcgagctttgc agacctgcgg tcagccctcc atgcgcaaga 1740 ctaaagcagc ggaagaggaggtgggcctct aggatctttg tcttctggct ggaggtgctt 1800 ttggaggttg ggtgctgggcattcagtcgc tcctctcacg cggctgcctt atcgggaagg 1860 aaatttgttt gccaaataaagactgacaca gaaaatcagg tcagtgtctc tgggctttgt 1920 gcaagctcag tttgaaaagggtttattccc atcactgccc aggacaccct gtggctttac 1980 ttgctcatgg tcagccaagcttacccttca cactgagaag tcatttctgg ctacttcctt 2040 gggctcagtt ccctgggtcatcagccatca aatcttgttg 2080 29 2128 DNA Homo sapiens misc_feature IncyteID No 1682513CB1 29 ctggccctag ggagctgccc ctgtcgctgg ctgcctgcaccaaccagccc cacattgtca 60 actacctgac ggagaacccc cacaagaagg cggacatgcggcgccaggac tcgcgaggca 120 acacagtgct gcatgcgctg gtggccattg ctgacaacacccgtgagaac accaagtttg 180 ttaccaagat gtacgacctg ctgctgctca agtgtgcccgcctcttcccc gacagcaacc 240 tggaggccgt gctcaacaac gacggcctct cgcccctcatgatggctgcc aagacgggca 300 agattgggaa ccgccacgag atgctggctg tggagcccatcaatgaactg ctgcgggaca 360 agtggcgcaa gttcggggcc gtctccttct acatcaacgtggtctcctac ctgtgtgcca 420 tggtcatctt cactctcacc gcctactacc agccgctggagggcacaccg ccgtaccctt 480 accgcaccac ggtggactac ctgcggctgg ctggcgaggtcattacgctc ttcactgggg 540 tcctgttctt cttcaccaac atcaaagact tgttcatgaagaaatgccct ggagtgaatt 600 ctctcttcat tgatggctcc ttccagctgc tctacttcatctactctgtc ctggtgatcg 660 tctcagcagc cctctacctg gcagggatcg aggcctacctggccgtgatg gtctttgccc 720 tggtcctggg ctggatgaat gccctttact tcacccgtgggctgaagctg acggggacct 780 atagcatcat gatccagaag attctcttca aggaccttttccgattcctg ctcgtctact 840 tgctcttcat gatcggctac gcttcagccc tggtctccctcctgaacccg tgtgccaaca 900 tgaaggtgtg caatggggac cagaccaact gcacagtgcccacttacccc tcgtgccgtg 960 acagcgagac cttcagcacc ttcctcctgg acctgtttaagctgaccatc ggcatgggcg 1020 acctggagat gctgagcagc accaagtacc ccgtggtcttcatcatcctg ctggtgacct 1080 acatcatcct cacctttgtg ctgctcctca acatgctcattgccctcatg ggcgagacag 1140 tgggccaggt ctccaaggag agcaagcaca tctggaagctgcagtgggcc accaccatcc 1200 tggacattga gcgctccttc cccgtattcc tgaggaagtccttccgctct ggggagatgg 1260 tcaccgtggg caagagctcg gacggcactc ctgaccgcaggtggtgcttc agggtggatg 1320 aggtgaactg gtctcactgg aaccagaact tgggcatcatcaacgaggac ccgggcaaga 1380 atgagaccta ccagtattat ggcttctcgc ataccgtgggccgcctccgc agggatcgct 1440 ggtcctcggt ggtaccccgc gtggtggaac tgaacaagaactcgaacccg gacgaggtgg 1500 tggtgcctct ggacagcacg gggaaccccc gctgcgatggccaccagcag ggttaccccc 1560 gcaagtggag gactgatgac gccccgctct agggactgcagcccagcccc agcttctctg 1620 cccactcatt tctagtccag ccgcatttca gcagtgccttctggggtgtc cccccacacc 1680 ctgctttggc cccagaggcg agggaccagt ggaggtgccagggaggcccc aggaccctgt 1740 ggtcccctgg ctctgcctcc ccaccctggg gtgggggctcccggccacct gtcttgctcc 1800 tatggagtca cataagccaa cgccagagcc cctccacctcaggccccagc ccctgcctct 1860 ccattattta tttgctctgc tctcaggaag cgacgtgacccctgccccag ctggaacctg 1920 gcagaggcct taggaccccg ttccaagtgc actgcccggccaagccccag cctcagcctg 1980 cgcctgagct gcatgcgcca ccatttttgg cagcgtggcagctttgcaag gggctggggc 2040 cctcggcgtg gggccatgcc ttctgtgtgt tctgtagtgtctgggatttg ccggtgctca 2100 ataaatgttt attcattgaa aaaaaaaa 2128 30 2825DNA Homo sapiens misc_feature Incyte ID No 2446438CB1 30 cgttgtgcacgtaattcggc tcgacgtgtg tccagatggt cagtctctgg tggctagcct 60 gtcctgacaggggagagtta agctcccgtt ctccaccgtg ccggctggcc aggtgggctg 120 agggtgaccgagagaccaga acctgcttgc tggagcttag tgctcagagc tggggaggga 180 ggttccgccgctcctctgct gtcagcgccg gcagcccctc ccggcttcac ttcctcccgc 240 agcccctgctactgagaagc tccgggatcc cagcagccgc cacgccctgg cctcagcctg 300 cggggctccagtcaggccaa caccgacgcg cagctgggag gaagacagga cccttgacat 360 ctccatctgcacagaggtcc tggctggacc gagcagcctc ctcctcctag gatgacctca 420 ccctccagctctccagtttt caggttggag acattagatg caggccaaga agatggctct 480 gaggcggacagaggaaagct ggattttggg agcgggctgc ctcccatgga gtcacagttc 540 cagggcgaggaccggaaatt cgcccctcag ataagagtca acctcaacta ccgaaaggga 600 acaggtgccagtcagccgga tccaaaccga tttgaccgag atcggctctt caatgcggtc 660 tcccggggtgtccccgagga tctggctgga cttccagagt acctgagcaa gaccagcaag 720 tacctcaccgactcggaata cacagagggc tccacaggta agacgtgcct gatgaaggct 780 gtgctgaaccttaaggacgg ggtcaatgcc tgcattctgc cactgctgca gatcgaccgg 840 gactctggcaatcctcagcc cctggtaaat gcccagtgca cagatgacta ttaccgaggc 900 cacagcgctctgcacatcgc cattgagaag aggagtctgc agtgtgtgaa gctcctggtg 960 gagaatggggccaatgtgca tgcccgggcc tgcggccgct tcttccagaa gggccaaggg 1020 acttgcttttatttcggtga gctacccctc tctttggccg cttgcaccaa gcagtgggat 1080 gtggtaagctacctcctgga gaacccacac cagcccgcca gcctgcaggc cactgactcc 1140 cagggcaacacagtcctgca tgccctagtg atgatctcgg acaactcagc tgagaacatt 1200 gcactggtgaccagcatgta tgatgggctc ctccaagctg gggcccgcct ctgccctacc 1260 gtgcagcttgaggacatccg caacctgcag gatctcacgc ctctgaagct ggccgccaag 1320 gagggcaagatcgagatttt caggcacatc ctgcagcggg agttttcagg actgagccac 1380 ctttcccgaaagttcaccga gtggtgctat gggcctgtcc gggtgtcgct gtatgacctg 1440 gcttctgtggacagctgtga ggagaactca gtgctggaga tcattgcctt tcattgcaag 1500 agcccgcaccgacaccgaat ggtcgttttg gagcccctga acaaactgct gcaggcgaaa 1560 tgggatctgctcatccccaa gttcttctta aacttcctgt gtaatctgat ctacatgttc 1620 atcttcaccgctgttgccta ccatcagcct accctgaaga agcaggccgc ccctcacctg 1680 aaagcggaggttggaaactc catgctgctg acgggccaca tccttatcct gctagggggg 1740 atctacctcctcgtgggcca gctgtggtac ttctggcggc gccacgtgtt catctggatc 1800 tcgttcatagacagctactt tgaaatcctc ttcctgttcc aggccctgct cacagtggtg 1860 tcccaggtgctgtgtttcct ggccatcgag tggtacctgc ccctgcttgt gtctgcgctg 1920 gtgctgggctggctgaacct gctttactat acacgtggct tccagcacac aggcatctac 1980 agtgtcatgatccagaaggt catcctgcgg gacctgctgc gcttccttct gatctactta 2040 gtcttccttttcggcttcgc tgtagccctg gtgagcctga gccaggaggc ttggcgcccc 2100 gaagctcctacaggccccaa tgccacagag tcagtgcagc ccatggaggg acaggaggac 2160 gagggcaacggggcccagta caggggtatc ctggaagcct ccttggagct cttcaaattc 2220 accatcggcatgggcgagct ggccttccag gagcagctgc acttccgcgg catggtgctg 2280 ctgctgctgctggcctacgt gctgctcacc tacatcctgc tgctcaacat gctcatcgcc 2340 ctcatgagcgagaccgtcaa cagtgtcgcc actgacagct ggagcatctg gaagctgcag 2400 aaagccatctctgtcctgga gatggagaat ggctattggt ggtgcaggaa gaagcagcgg 2460 gcaggtgtgatgctgaccgt tggcactaag ccagatggca gccccgatga gcgctggtgc 2520 ttcagggtggaggaggtgaa ctgggcttca tgggagcaga cgctgcctac gctgtgtgag 2580 gacccgtcaggggcaggtgt ccctcgaact ctcgagaacc ctgtcctggc ttcccctccc 2640 aaggaggatgaggatggtgc ctctgaggaa aactatgtgc ccgtccagct cctccagtcc 2700 aactgatggcccagatgcag caggaggcca gaggacagag cagaggatct ttccaaccac 2760 atctgctggctctggggtcc cagtgaattc tggtggcaaa tatatatttt cactaaaaaa 2820 aaaaa 282531 1718 DNA Homo sapiens misc_feature Incyte ID No 2817822CB1 31gcctcggtgt tcccacctag gggcgggcag ccaggggcac ttccgctggc ccaagtgatc 60tgcatgtggc agggctgcgc agtggagcgg ccagtgggca ggatgacgag ccagacccct 120ctgccccagt ccccccggcc caggcggcca acgatgtcta ctgttgtgga gctgaacgtc 180gggggtgagt tccacaccac caccctgggt accctgagga agtttccggg ctcaaagctg 240gcagagatgt tctctagctt agccaaggcc tccacggacg cggagggccg cttcttcatc 300gaccgcccca gcacctattt cagacccatc ctggactacc tgcgcactgg gcaagtgccc 360acacagcaca tccctgaagt gtaccgtgag gctcagttct acgaaatcaa gcctttggtc 420aagctgctgg aggacatgcc acagatcttt ggtgagcagg tgtctcggaa gcagtttttg 480ctgcaagtgc cgggctacag cgagaacctg gagctcatgg tgcgcctggc acgtgcagaa 540gccataacag cacggaagtc cagcgtgctt gtgtgcctgg tggaaactga ggagcaggat 600gcatattatt cagaggtcct gtgttttctg caggataaga agatgttcaa gtctgttgtc 660aagtttgggc cctggaaggc ggtcctagac aacagcgacc tcatgcactg cctggagatg 720gacattaagg cccaggggta caaggtattc tccaagttct acctgacgta ccccaccaaa 780agaaacgaat tccattttaa catttattca ttcaccttca cctggtggtg atcctcagga 840gcagagactg ttatgaattc tggcgtggct tatgaaatta aaagttgcca tcaaagccat 900tttcttttaa tttcacaaac atcaggcaat ttccagggtt ggtctagagt cttgccacta 960aatattgatc actcgtttaa ggactttcca ctccattgca actgatgcca ctatatttgc 1020ctagcaactt gcagctactt ccttttcaaa gcctcatgta tctcccagac ccttctcttg 1080aagtccaata acaagaccaa gtaagaatgt ttcaacaatg cgttggcaag agatgtgaga 1140tgacaacagg aacatacaag atactgtgaa tctagatgtt ctgacctaaa gatgtagtct 1200acatagcccc agcttggggt ccaatccatc tgtccctggc atgtgccttc atgtagtagg 1260tgctttcctg atcccctttg cgagatgctg tgggtgctaa cacctcagag ctgtcctctt 1320ctctagagtg gaggttttca aagtgcatca tcagcattac ctgtgaactt gctggaaata 1380caaatcctca ggccccacct cagacctact gaatcagaat ctctgggggt tggcacagca 1440ttctgattta ccaaaccctc caagtgattt tgatgtattc taattttgag accatctcta 1500gaaaagaatt gctacctctt gtatggaggt acaaaagact gacctcttac atcaaggaac 1560ttcctttccc agagctcctc atggaatcaa gctgaagtca gtcttcttct gagagcacat 1620tcttactcag tttttttcct ctgtcctacg ctgcttccct cactcccctt ctcctaagag 1680cactccatca ataaaccact tgcacgagaa aaaaaaaa 1718 32 2000 DNA Homo sapiensmisc_feature Incyte ID No 4009329CB1 32 gacgaatttg aaaccagggg gtgtcctgtttgaacttggt gccagataga gtaactcgga 60 ctccagttgg aggggttcgg gagaaccatagaagaggaag ggccgtgtct tccgtggaca 120 ggccaccgga gccgccagct gtttggaactgagctactgc agaaagggaa gtggagagta 180 agggccaggc cccgtggggg cagatggccggcagaaggct gaatctgcgc tgggcactga 240 gtgtgctttg tgtgctgcta atggcggagacagtgtctgg gactaggggc tcgtctacag 300 gagctcacat tagcccccag tttccagcttcaggtgtgaa ccagaccccc gtggtagact 360 gccgcaaggt gtgtggcctg aatgtctctgaccgctgtga cttcatccgg accaaccctg 420 actgccacag tgatgggggg tacctggactacctggaagg catcttctgc cacttccctc 480 ccagcctcct ccctctggct gtcactctctacgtttcctg gctgctctac ctgtttctga 540 ttctgggagt caccgcagcc aagtttttctgccccaactt gtcggccatt tctaccacac 600 tgaagctctc ccacaacgtg gcaggcgtcaccttcctggc atttgggaat ggtgcacctg 660 acatcttcag tgccctggtg gccttctctgacccgcacac agccggcctg gcccttgggg 720 cactgtttgg cgctggcgtg ctggttaccacagtggtggc cggaggcatt accatcctac 780 accccttcat ggctgcctcc aggcccttcttcagggacat cgttttctac atggtggctg 840 tgttcctgac cttcctcatg ctcttccgtggcagggtcac cctggcatgg gctctgggtt 900 acctgggctt gtatgtgttc tatgtggtcactgtgattct ctgcacctgg atctaccaac 960 ggcaacggag aggatctctg ttctgccccatgccagttac tccagagatc ctctcagact 1020 ccgaggagga ccgggtatct tctaataccaacagctatga ctacggtgat gagtaccggc 1080 cgctgttctt ctaccaggag accacggctcagatcctggt ccgggccctc aatcccctgg 1140 attacatgaa gtggagaagg aaatcagcatactggaaagc cctcaaggtg ttcaagctgc 1200 ctgtggagtt cctgctgctc ctcacagtccccgtcgtgga cccggacaag gatgaccaga 1260 actggaaacg gcccctcaac tgtctgcatctggttatcag ccccctggtt gtggtcctga 1320 ccctgcagtc ggggacctat ggtgtctatgagataggcgg cctcgttccc gtctgggtcg 1380 tggtggtgat cgcaggcaca gccttggcttcagtgacctt ttttgccaca tctgacagcc 1440 agccccccag gcttcactgg ctctttgctttcctgggctt tctgaccagc gccctgtgga 1500 tcaacgcggc cgccacagag gtggtgaacatcttgcggtc cctgggtgtg gtcttccggc 1560 tgagcaacac tgtgctgggg ctcacgctgctggcctgggg gaacagcatt ggagatgcct 1620 tctcggattt cacactggct cgccagggctacccacggat ggcgttctcc gcctgctttg 1680 gcggcatcat cttcaacatc ctcgtgggtgtggggctggg ctgcctgctc cagatctccc 1740 gaagccacac agaagtgaag ctggagccagacggactgct ggtgtgggtc ctggcaggcg 1800 ccctggggct cagcctcgtc ttctccctggtctcagtccc attgcagtgc ttccagctca 1860 gcagagtcta tggcttctgc ctgctcctcttctacctgaa cttccttgtc gtggccctcc 1920 tcattgaatt tggagtgatt cacctgaaaagcatgtgact gaagccgctt agtgctgtgg 1980 cctcactgca ggcaggagcc 2000 33 2216DNA Homo sapiens misc_feature Incyte ID No 6618083CB1 33 gaaaactcttcctgaaggag atgcagagga agattcgaac tggaggaaaa ccctaaaata 60 aacaataacaacaaaagttc aaaacctgaa aagtgaacca tgaagctcag taaaaaggac 120 cgaggagaagatgaagaaag tgattcagcg aaaaagaaat tggactggtc ctgctcgctc 180 ctcgtggcctccctcgcggg cgccttcggc tcctccttcc tctacggcta caacctgtcg 240 gtggtgaatgcccccacccc gtacatcaag gccttttaca atgagtcatg ggaaagaagg 300 catggacgtccaatagaccc agacactctg actttgctct ggtctgtgac tgtgtccata 360 ttcgccatcggtggacttgt ggggacgtta attgtgaaga tgattggaaa ggttcttggg 420 aggaagcacactttgctggc caataatggg tttgcaattt ctgctgcatt gctgatggcc 480 tgctcgctccaggcaggagc ctttgaaatg ctcatcgtgg gacgcttcat catgggcata 540 gatggaggcgtcgccctcag tgtgctcccc atgtacctca gtgagatctc acccaaggag 600 atccgtggctctctggggca ggtgactgcc atctttatct gcattggcgt gttcactggg 660 cagcttctgggcctgcccga gctgctggga aaggagagta cctggccata cctgtttgga 720 gtgattgtggtccctgccgt tgtccagctg ctgagccttc cctttctccc ggacagccca 780 cgctacctgctcttggagaa gcacaacgag gcaagagctg tgaaagcctt ccaaacgttc 840 ttgggtaaagcagacgtttc ccaagaggta gaggaggtcc tggctgagag ccacgtgcag 900 aggagcatccgcctggtgtc cgtgctggag ctgctgagag ctccctacgt ccgctggcag 960 gtggtcaccgtgattgtcac catggcctgc taccagctct gtggcctcaa tgcaatttgg 1020 ttctataccaacagcatctt tggaaaagct gggatccctc tggcaaagat cccatacgtc 1080 accttgagtacagggggcat cgagactttg gctgccgtct tctctggttt ggtcattgag 1140 cacctgggacggagacccct cctcattggt ggctttgggc tcatgggcct cttctttggg 1200 accctcaccatcacgctgac cctgcaggac cacgccccct gggtccccta cctgagtatc 1260 gtgggcattctggccatcat cgcctctttc tgcagtgggc cagctgtttt cccagaagaa 1320 acggtaaatgtcagcattgt atctgagtga aaagttgacc ttcttcccca cccatgcaca 1380 caaacaagccagattggact catctgcata tctgcctgaa gttctttgct aaccaaaaat 1440 cactaagcttagccttctct gttttttttt tcctaagccc tcccaagact ttttgcaatg 1500 atcctgattctgttccaagt gtttgcaact gtggctttct tttgactgta gaacatgctg 1560 catttccagggctttaaatg ctgggctccc catcagtgtc tatgggactc cctggaggga 1620 aggccacctgcacctcccaa tcccagatca cctgtcagcc cctgccctcc gcttcctcaa 1680 tccatcttcaaccccctgtg ttgacccagc acctgggcct tgctggctag caatgacttt 1740 agccacaagatggaccaggg tttagaagct tcatttaaac tcacattgac agtgtacagt 1800 ttaaagcctcagggaactta cctgtctaag aaaagctgcc acttagacca tgagaccatc 1860 ttgcatcttcctaagtggac agggaagagc aagtccccag gggagccacc cgggaaagtg 1920 tggcaggaagatgctcagag ctgaatggca gagagactca tgggcctgct ctccatgatt 1980 aaagaagagggatggatctc ccaggagagg gccaggaggc cgcctgaggc agcttctgtg 2040 aggaacaggtcgatgtaaga agacttgaca aggagttgaa attaggtgaa agcaaagaaa 2100 gaaaacaagagaggcagttt cctgctgcat attttatttg tgtgcataac cccaaggcag 2160 tggcagggaagtctaataaa tgaggcaaaa taaaagagct tcacctttta aaaaaa 2216 34 1995 DNA Homosapiens misc_feature Incyte ID No 7472002CB1 34 atgaccgaaa aaaccaatggtgtgaagagc tccccagcca ataatcacaa ccatcatgca 60 cctcctgcca tcaaggccaatggcaaagat gaccacagga caagcagcag gccacactct 120 gcagctgacg atgacacctcctcagaactg cagaggctgg cagacgtgga tgccccacag 180 cagggaagga gtggcttccgcaggatagtt cgcctggtgg ggatcatcag agaatgggcc 240 aacaagaatt tccgagaggaggaacctagg cctgactcat tcctcgagcg ttttcgtggg 300 cctgaactcc agactgtgaccacacaggag ggggatggca aaggcgacaa ggatggcgag 360 gacaaaggca ccaagaagaaatttgaacta tttgtcttgg acccagctgg ggattggtac 420 tactgctggc tatttgtcattgccatgccc gtcctttaca actggtgcct gctggtggcc 480 agagcctgct tcagtgacctacagaaaggc tactacctgg tgtggctggt gctggattat 540 gtctcagatg tggtctacattgcggacctc ttcatccgat tgcgcacagg tttcctggag 600 caggggctgc tggtcaaagataccaagaaa ctgcgagaca actacatcca caccctgcag 660 ttcaagctgg atgtggcttccatcatcccc actgacctga tctattttgc tgtggacatc 720 cacagccctg aggtgcgcttcaaccgcctg ctgcactttg cccgcatgtt tgagttcttt 780 gaccggacag agacacgcaccaactaccct aacatcttcc gcatcagcaa ccttgtcctc 840 tacatcttgg tcatcatccactggaatgcc tgcatctatt atgccatctc caaatccata 900 ggctttgggg tcgacacctgggtttaccca aacatcactg accctgagta tggctacctg 960 gctagggaat acatctattgcctttactgg tccacactga ctctcactac cattggggag 1020 acaccacccc ctgtaaaggatgaggagtac ctatttgtca tctttgactt cctgattggc 1080 gtcctcatct ttgccaccatcgtgggaaat gtgggctcca tgatctccaa catgaatgcc 1140 acccgggcag agttccaggctaagatcgat gccgtgaaac actacatgca gttccgaaag 1200 gtcagcaagg ggatggaagccaaggtcatt aggtggtttg actacttgtg gaccaataag 1260 aagacagtgg atgagcgagaaattctcaag aatctgccag ccaagctcag ggctgagata 1320 gccatcaatg tccacttgtccacactcaag aaagtgcgca tcttccatga ttgtgaggct 1380 ggcctgctgg tagagctggtactgaaactc cgtcctcagg tcttcagtcc tggggattac 1440 atttgccgca aaggggacatcggcaaggag atgtacatca ttaaggaggg caaactggca 1500 gtggtggctg atgatggtgtgactcagtat gctctgctgt cggctggaag ctgctttggc 1560 gagatcagta tccttaacattaagggcagt aaaatgggca atcgacgcac agctaatatc 1620 cgcagcctgg gctactcagatctcttctgc ttgtccaagg atgatcttat ggaagctgtg 1680 actgagtacc ctgatgccaagaaagtccta gaagagaggg gtcgggagat cctcatgaag 1740 gagggactgc tggatgagaacgaagtggca accagcatgg aggtcgacgt gcaggagaag 1800 ctagggcagc tggagaccaacatggaaacc ttgtacactc gctttggccg cctgctggct 1860 gagtacacgg gggcccagcagaagctcaag cagcgcatca cagttctgga aaccaagatg 1920 aaacagaaca atgaagatgactacctgtct gatgggatga acagccctga gctggctgct 1980 gctgacgagc cataa 199535 988 DNA Homo sapiens misc_feature Incyte ID No 1812692CB1 35cttgggtgaa agaaaatcct gcttgacaaa aaccgtcact taggaaaaga tgtcctttcg 60ggcagccagg ctcagcatga ggaacagaag gaatgacact ctggacagca cccggaccct 120gtactccagc gcgtctcgga gcacagactt gtcttacagt gaaagcgact tggtgaattt 180tattcaagca aattttaaga aacgagaatg tgtcttcttt accaaagatt ccaaggccac 240ggagaatgtg tgcaagtgtg gctatgccca gagccagcac atggaaggca cccagatcaa 300ccaaagtgag aaatggaact acaagaaaca caccaaggaa tttcctaccg acgcctttgg 360ggatattcag tttgagacac tggggaagaa agggaagtat atacgtctgt cctgcgacac 420ggacgcggaa atcctttacg agctgctgac ccagcactgg cacctgaaaa cacccaacct 480ggtcatttct gtgaccgggg gcgccaagaa cttcgccctg aagccgcgca tgcgcaagat 540cttcagccgg ctcatctaca tcgcgcagtc caaaggtgct tggattctca cgggaggcac 600ccattatggc ctgatgaagt acatcgggga ggtggtgaga gataacacca tcagcaggag 660ttcagaggag aatattgtgg ccattggcat agcagcttgg ggcatggtct ccaaccggga 720caccctcatc aggaattgcg atgctgaggt accggtggga caggaggagg tctgctaggt 780cacatggaag aaagaccatg gcatgggcct gtggcctgaa ccctggggct ctgtgatgga 840gccagccaga tcatggggaa gtctgccttt caaggagtgc ctttgggacc ttaaaggaat 900tgaaaacaag gatgacgtac ctaattaact gctgggaaag agttaacaat gaatgttttg 960ttcattaaaa tgtgttctca gcaatctc 988 36 3179 DNA Homo sapiens misc_featureIncyte ID No 3232992CB1 36 gcggagcggc ggcgccggcg ccggggggcg cagcgaggggctggcggtag cggttgctgc 60 ggggcgcggg gcgcgggcgg cgctggagtc tcggccgcgggcgatgaggt gcagacgctg 120 tcgggcagcg taaggcgggc cccgaccgga ccccccggcacccccggcac ccccggctgc 180 gcagctactg caaaggggcc ccggcgctca gcagcccaaaccggccagct tgggccgcgg 240 gcggggggca agccgccgcc atcctcagct tgggcaacgtgctcaactac ctggacaggt 300 acaccgtggc aggcgtcctt ctggacatcc agcagcactttggggtcaag gaccgaggcg 360 ccggcctgct gcagtcagtg ttcatctgta gcttcatggtggctgccccc atcttcggct 420 acctgggcga ccgcttcaac aggaaggtga ttctcagctgcggcattttc ttctggtcgg 480 ccgtcacctt ctccagctcc ttcattcccc agcagtacttctggctgctg gtcctgtccc 540 gggggctggt gggcatcggg gaggccagct actccaccatcgcccccact atcattggcg 600 acctcttcac caagaacacg cgtacgctca tgctgtccgtcttctacttc gccatcccac 660 tgggcagtgg cctgggctac attactggct ccagcgtgaagcaggcagcc ggagactggc 720 actgggcatt gcgggtgtcc cctgtcctgg gcatgatcacaggaacactc atcctcattc 780 tggtcccagc cactaaaagg ggtcatgccg accagctcggggaccagctc aaggcccgga 840 cctcatggct ccgagatatg aaggccctga ttcgaaaccgcagctacgtc ttctcctccc 900 tggccacgtc ggctgtctcc ttcgccacgg gggccctgggcatgtggatc ccgctctacc 960 tgcaccgcgc ccaagttgtg cagaagacag cagagacgtgcaacagcccg ccctgtgggg 1020 ccaaggacag cctcatcttt ggggccatca cctgctttacgggatttctg ggcgtggtca 1080 cgggggcagg agccacgcgc tggtgccgcc tgaagacccagcgggccgac ccactggtgt 1140 gtgccgtggg catgctgggc tctgccatct tcatctgcctgatcttcgtg gctgccaaga 1200 gcagcatcgt aggagcctat atctgtatct tcgtcggggagacgctgctg ttttctaact 1260 gggccatcac tgcagacatc ctcatgtacg tggtcatccccacgcggcgc gccactgccg 1320 tggccttgca gagcttcacc tcccacctgc tgggggacgccgggagcccc tacctcattg 1380 gctttatctc agacctgatc cgccagagca ctaaggactccccgctctgg gagttcctga 1440 gcctgggcta cgcgctcatg ctctgccctt tcgtcgtggtcctgggcggc atgttcttcc 1500 tcgccactgc gctcttcttc gtcagcgacc gcgccagggctgagcagcag gtgaaccagc 1560 tggcgatgcc gcccgcatct gtgaaagtct gaggtggtgccattgggaca atgaagaacc 1620 cacactccca cctcgtctgg gaggtgtcct acagcgtccgggaccggctg ggctgcccca 1680 aagctttctg tgtgatccac ggctaggcac ccaccctctctggcccaggc ctgctgagtg 1740 gccctggcat caagaggagg ctgtgtcctc agttaccctggaaggatgtg tgtgttggag 1800 ccacacggtt ggacaggttc ccagccctag gtttgggccgcagggcccct ggggccaagg 1860 aagaagacag ccccaagtgg gtgtccgggg agagcctggcctgccaccag cttatgtgat 1920 cttgggcaag tccctgccct ccctggaacg aagggccagggggctggact ttcccacaca 1980 acttgctggg caaagcacga tctgcagctt tgaagactcaacagaccctg gaccatacgg 2040 agagcaggtg gcccaggcct cagggcggca gtcccggctttgaggctcac gcgagggcct 2100 ggtatgcagg gaccactgct cagctgggcc tcggaccttggggatattgg acgcaacctg 2160 gcaaatgaag ctgggcgccc aagtctctgg gtactccctggaggacactg tctcactgtc 2220 tcgggttggc tcccagcctg gaggtcccag atggggactgttctgacaag ctggcatcac 2280 caggggtgaa ggccctggct gcagctgtac accacctgtgcccccaggct caaggtctct 2340 ggcaggtgca caccagccca actctgcagg gcttctctccctgccaccac cccccaagcc 2400 aggaccccac tccttccccg aggctgagct gagccttttccaggggcagg gcccaggaga 2460 ccattcccag aatccatggg gcagtagcca gggctccggctgctggagga agcagctatc 2520 cacaaagctt cctgccccag agctgaggct gaggccccgggagaggcggc ccctacccaa 2580 acactggctg ctggcattcc accaagtgac cccaggggccaggccttcga tcacccacct 2640 cccatccatg cacacaccag gatgcagctg ccaacttcacaccagcccca acccgctttg 2700 ggggagctta gccccctgcg tcacccactg cctgcacttctgctgcaatc aaggtggttc 2760 tggtgcgggg gtggggtggg gggtgaggcc ttgtggccaatgggggaccc cccaagagcc 2820 agcttggaca atgctcttct tgccccttag ttactggctggctgtggctt cagtggtgtg 2880 taagcaggtg gaatactcac ccaccaagct ctggggtaccccgagggcct gacaagagga 2940 tggggtgggg gtggcatcct ccaaagacca gcctccacccccactccagc ctcagcgggg 3000 ccccagcgat gttttcttgt tgtacaagaa ccaggtccgagtgttgcctc ctcttccttc 3060 cggaagccaa actgctcctt tattttttag agctgctgattgtgaatctc agagtcttaa 3120 gagagaagcc aaatatattc ctcttgtaaa tgaagaaataaacctattta aatcacaaa 3179 37 1986 DNA Homo sapiens misc_feature IncyteID No 3358383CB1 37 ggagtatctg agcaaattat ttcttacgtg actttagagaaaacggctac ctatctgacc 60 ccaaaacgac ttgaggaaac tgtttccacg gtcctgctgcaggggggaag cacagtcgtc 120 aagaagagag tggggtcagg atcaaaacac atttagtgtgacttagggaa agaaaacatt 180 ttccctcttt gaacctctct ggatacagtc attttgcctctacttgagga tcaactgttc 240 aacctcaatg gcctttcagg acctcctggg tcacgctggtgacctgtgga gattccagat 300 ccttcagact gtttttctct caatctttgc tgttgctacataccttcatt ttatgctgga 360 gaacttcact gcattcatac ctggccatcg ctgctgggtccacatcctgg acaatgacac 420 tgtctctgac aatgacactg gggccctcag ccaagatgcactcttgagaa tctccatccc 480 actggactca aacatgaggc cagagaagtg tcgtcgctttgttcatcctc agtggcagct 540 ccttcacctg aatgggacct tccccaacac aagtgacgcagacatggagc cctgtgtgga 600 tggctgggtg tatgacagaa tctccttctc atccaccatcgtgactgagt gggatctggt 660 atgtgactct caatcactga cttcagtggc taaatttgtattcatggctg gaatgatggt 720 gggaggcatc ctaggcggtc atttatcaga caggtttgggagaaggttcg tgctcagatg 780 gtgttacctc caggttgcca ttgttggcac ctgtgcagccttggctccca ccttcctcat 840 ttactgctca ctacgcttct tgtctgggat tgctgcaatgagcctcataa caaatactat 900 tatgttaata gccgagtggg caacacacag attccaggccatgggaatta cattgggaat 960 gtgcccttct ggtattgcat ttatgaccct ggcaggcctggcttttgcca ttcgagactg 1020 gcatatcctc cagctggtgg tgtctgtacc atactttgtgatctttctga cctcaagttg 1080 gctgctagag tctgctcggt ggctcattat caacaataaaccagaggaag gcttaaagga 1140 acttagaaaa gctgcacaca ggagtggaat gaagaatgccagagacaccc taaccctgga 1200 gattttgaaa tccaccatga aaaaagaact ggaggcagcacaaaaaaaaa aaccttctct 1260 gtgtgaaatg ctccacatgc ccaacatatg taaaaggatctccctcctgt cctttacgag 1320 atttgcaaac tttatggcct attttggcct taatctccatgtccagcatc tggggaacaa 1380 tgttttcctg ttgcagactc tctttggtgc agtcatcctcctggccaact gtgttgcacc 1440 ttgggcactg aaatacatga cccgtcgagc aagccagatgcgtctcatgt acctactggc 1500 aatctgcttt atggccatca tatttgtgcc acaagaaatgcagacgctgc gtgaggtttt 1560 ggcaacactg ggcttaggag cgtcggctct gaccaatacccttgcttttg cccatggaaa 1620 tgaagtaatt cccaccataa tcagggcaag agctatggggatcaatgcaa cctttgctaa 1680 tatagcagga gccctggctc ccctcatgat gatcctaagtgtgtattctc cacccctgcc 1740 ctggatcatc tatggagtct tccccttcat ctctggctttgctttcctcc tccttcctga 1800 aaccaggaac aagcctctgt ttgacaccat ccaggatgagaaaaatgaga gaaaagaccc 1860 cagagaacca aagcaagagg atccgagagt ggaagtgacgcagttttaag gaattccagg 1920 agctgactgc cgatcaatga gccagatgaa gggaacaatcaggactattc ctagacacta 1980 gcaaat 1986 38 3294 DNA Homo sapiensmisc_feature Incyte ID No 4250091CB1 38 tgtaagacag gaaagggatc tatttgatgtctatcttcag atatattggc agttttcctt 60 aagctattta gttcctcatc tgttgctttttcattttgta tactgcaagt tcccaggcaa 120 ctcgaatttg caaacacagc catggatacactatttacct tacagtagtt tcctgggaat 180 ctaagtctgg tttttgttat tcttccctcccctccactgc ataatcatgt ataactagca 240 acatttatgg ttataggttg atttcctaagtgtggctgat ggtagcctct agtttgaagt 300 gagggaagaa tgagtagtca ggaactggtcactttgaatg tgggagggaa gatattcacg 360 acaaggtttt ctacgataaa gcagtttcctgcttctcgtt tggcacgcat gttagatggc 420 agagaccaag aattcaagat ggttggtggccagatttttg tagacagaga tggtgatttg 480 tttagtttca tcttagattt tttgagaactcaccagcttt tattacccac tgaattttca 540 gactatctta ggcttcagag agaggctcttttctatgaac ttcgttctct agttgatctc 600 ttaaacccat acctgctaca gccaagacctgctcttgtgg aggtacattt cctaagccgg 660 aacactcaag cttttttcag ggtgtttggctcttgcagca aaacaattga gatgctaaca 720 gggaggatta cagtgtttac agaacaaccttcggcgccga cctggaatgg taactttttc 780 cctcctcaga tgaccttact tccactgcctccacaaagac cttcttacca tgacctggtt 840 ttccagtgtg gttctgacag cactactgataaccaaactg gagtcaggta ttttgtactt 900 tgcagtattt ctcttgtata ccagtttgtgatgttttctc taaaaacttg aagttcctca 960 ggcctgtaac ttctggaaaa gatgattattcaaaataatg ttttggggta accagtggag 1020 ttgggtagaa tgaccaaata attattttccaaactgggat actttttaga gtgaaagggg 1080 ctattattag gtgggacaaa aggaataaatgaagactgcc cagaaaaaac tgagactatg 1140 gacattcaaa tcatgggaga aaataattttgtagattatg ttccattgct aatgaatttg 1200 acttagaaaa gaattgcctt atttttaagagattgtttca gtggttcaca taaaggctcg 1260 ctcactggtt tctcttgagt tccttacacactatataagt tgttctttca gttttatgat 1320 tcaactactg tttttccttc agctgactttatttttaaac acccttaaag acagatatat 1380 ctcatggcaa atttggtatc ctgttacagccttggctctt aaacaactca aaatattggg 1440 ataggctgtc agtatgttaa ggatagttgctcctgagtca attcttcact tactccctct 1500 gttgttcttg gctggatcct aacgctgatttccactctgc tgtcacaaac atttttcccc 1560 ccgtaaaatg tcttaatgct gtcctaccattattttacca actgtgaaag ctggctttaa 1620 tttttaggag gaaaagaaaa gcctgcatgtgttctttatt ggtatcattt aaaatatact 1680 tttttttttt ttttggtaaa ggtaggcgtattttaagata ttttcttaac ttgagcagta 1740 gccaacagga aggataccag tgtctctctctcttagcgac acactccttg gtcttgctta 1800 ccaactggag gacactaggt agaataaccgagtatgacaa ttcttaattg tttacatttt 1860 ataacttcct gtccttcaaa agagtttgaaatgtcatttt gggaaaagag agccagtcaa 1920 gctagtaggc tgattgtgaa gaaaatctaataccttatct ttatctcaaa cctctgtaca 1980 actttatttt cattgatggg atactttaacaaaaatgaaa ttttttttgg tttttaaaat 2040 atgagtgatt atgacctctt tggggatcatgcttcaaaaa gtcagaaacc tagagacaaa 2100 actgtcattg atttttaaga agaaacacactaggtcaaaa gaagatgtcc tggaaatacg 2160 aagtactctt taaaaaccat gcatttggagaaagtaattg tttccttgaa aaacatgatt 2220 aaaaactaaa actgggatgt tcctgtgtgtacacagtgcc aaatggtttt ccctttttat 2280 gttgtgtttt agaaacagca cgaaagttttttccatttta aagtgagaaa acattatatt 2340 tagacttcca taattccaaa atcagaagctatttttaaaa ttagcatttt cttgcatcac 2400 caaatggtat tcaattgttt gaagctcaaaatttttacca ttccataaat gtttgtgaat 2460 ttttagacag tgccaattta aaagtagagatagccaatct gaatacggtg aaattatggg 2520 gatctctggt gattgggatg aaaactctggccttaaaagg tccactttta gtatataatt 2580 gcctaattag caatcatttt tattttttgctcactccctg gtctgaatct atctgtctat 2640 tcagatattt tttggtaggt ttggaaaatggagaagtgag cctaattggt gcctaattgt 2700 ctggtgtatc attcacttta ttcagtttgttctatcaata tgatttaccc ctcaaggtta 2760 acctagcagg ttgctcagtt attatctctcaaggtcacag tactagaaat acttggcttg 2820 catctttcag atgccattca tgttatcaagctcaaattat agttggtcac aggattctaa 2880 agtctttatt tgacttctcc tttttgaactggctcaaatg gaaaagtgta gttgctttta 2940 aatgttaaaa ataagtttaa actttatatttcccattggt ttcccctatt ttgtcctttc 3000 tttgtgtgct tgaaatattt tatttttcagtttgtcctca tagggaatca agtattttag 3060 ctaggtgatg tcttgcaagt acgttccactttgttacaat ctactatctg tatatactat 3120 ttgtatctta attcttttat gagatgttctgtaacatttt tctcactttg acaaatgttt 3180 ttagactgta cagtcaagat ctggcgcttgggggtaagtg gaatgatttg ctaatattga 3240 gaatctgttg tatcaaacat aataaactttttttgagatg tgaaaaaaaa aaaa 3294 39 2043 DNA Homo sapiens misc_featureIncyte ID No 70064803CB1 39 gcaacatggc ggctgccgtg gtgcagcgcc cgggctgagcgacagcaagt gcagcgggct 60 cctaccccgg gtgaggggtg gcctccgcgt gggatcgtgccctcttcagc ccgctcctgt 120 ccccgacatc acgtgtattc cgcacgtccc ctccgcgctgtgtgtctact gagacgggga 180 ggcgtgacag ggcccgggtc ccttctcagt ggtgctctgtgcttcagggc aagctccccg 240 tctccgggcg cacttccctc gcctgtgttc ggtccatcctcctttctcca gcctcctccc 300 ctcgcaggtg ggatcgtcgg tgggaccgga gcgcgggcgggcgcggcccc ccgggaccat 360 ggccgggtcc gacaccgcgc ccttcctcag ccaggcggatgacccggacg acgggccagt 420 gcctggcacc ccggggttgc cagggtccac ggggaacccgaagtccgagg agcccgaggt 480 cccggaccag gaggggctgc agcgcatcac cggcctgtctcccggccgtt cggctctcat 540 agtggcggtg ctgtgctaca tcaatctcct gaactacatggaccgcttca ccgtggctgg 600 cgtccttccc gacatcgagc agttcttcaa catcggggacagtagctctg ggctcatcca 660 gaccgtgttc atctccagtt acatggtgtt ggcacctgtgtttggctacc tgggtgacag 720 gtacaatcgg aagtatctca tgtgcggggg cattgccttctggtccctgg tgacactggg 780 gtcatccttc atccccggag agcatttctg gctgctcctcctgacccggg gcctggtggg 840 ggtcggggag gccagttatt ccaccatcgc gcccactctcattgccgacc tctttgtggc 900 cgaccagcgg agccggatgc tcagcatctt ctactttgccattccggtgg gcagtggtct 960 gggctacatt gcaggctcca aagtgaagga tatggctggagactggcact gggctctgag 1020 ggtgacaccg ggtctaggag tggtggccgt tctgctgctgttcctggtag tgcgggagcc 1080 gccaagggga gccgtggagc gccactcaga tttgccacccctgaacccca cctcgtggtg 1140 ggcagatctg agggctctgg caagaaatct catctttggactcatcacct gcctgaccgg 1200 agtcctgggt gtgggcctgg gtgtggagat cagccgccggctccgccact ccaacccccg 1260 ggctgatccc ctggtctgtg ccactggcct cctgggctctgcacccttcc tcttcctgtc 1320 ccttgcctgc gcccgtggta gcatcgtggc cacttatattttcatcttca ttggagagac 1380 cctcctgtcc atgaactggg ccatcgtggc cgacattctgctgtacgtgg tgatccctac 1440 ccgacgctcc accgccgagg ccttccagat cgtgctgtcccacctgctgg gtgatgctgg 1500 gagcccctac ctcattggcc tgatctctga ccgcctgcgccggaactggc ccccctcctt 1560 cttgtccgag ttccgggctc tgcagttctc gctcatgctctgcgcgtttg ttggggcact 1620 gggcggcgca gccttcctgg gcaccgccat cttcattgaggccgaccgcc ggcgggcaca 1680 gctgcacgtg cagggcctgc tgcacgaagc agggtccacagacgaccgga ttgtggtgcc 1740 ccagcggggc cgctccaccc gcgtgcccgt ggccagtgtgctcatctgag aggctgccgc 1800 tcacctacct gcacatctgc cacagctggc cctgggcccaccccacgaag ggcctgggcc 1860 taaccccttg gcctggccca gcttccagag ggaccctgggccgtgtgcca gctcccagac 1920 actacatggg tagctcaggg gaggaggtgg gggtccaggagggggatccc tctccacagg 1980 ggcagcccca agggctcggt gctatttgta acggaataaaatttgtagcc agacaaaaaa 2040 aaa 2043 40 1915 DNA Homo sapiensmisc_feature Incyte ID No 70356768CB1 40 caccactggg cgctgcgcgctgcccttccc tccgcgcaca ggctgccggc tcaccgcttg 60 ctaatggcag ccggggtctccctgggacag caagacctcc gctcaggccc ctctttcgaa 120 tgctccacgc cctcctgcgatctagaatga ttcagggcag gatcctgctc ctgaccatct 180 gcgctgccgg cattggtgggacttttcagt ttggctataa cctctctatc atcaatgccc 240 cgaccttgca cattcaggaattcaccaatg agacatggca ggcgcgtact ggagagccac 300 tgcccgatca cctagtcctgcttatgtggt ccctcatcgt gtctctgtat cccctgggag 360 gcctctttgg agcactgcttgcaggtccct tggccatcac gctgggaagg aagaagtccc 420 tcctggtgaa taacatctttgtggtgtcag cagcaatcct gtttggattc agccgcaaag 480 caggctcctt tgagatgatcatgctgggaa gactgctcgt gggagtcaat gcaggtgtga 540 gcatgaacat ccagcccatgtacctggggg agagcgcccc taaggagctc cgaggagctg 600 tggccatgag ctcagccatctttacggctc tggggatcgt gatgggacag gtggtcggac 660 tcagggagct cctaggtggccctcaggcct ggcccctgct gctggccagc tgcctggtgc 720 ccggggcgct ccagctcgcctccctgcctc tgctccctga aagcccgcgc tacctcctca 780 ttgactgtgg agacaccgaggcctgcctgg cagagacggg ttctcgcttg tccaggctgg 840 agtgctgtgg ctgttcataggcatgacccc attgttgatc agcacggaag ttttcttctt 900 ttttgttttt gtttttttggttttgtttgg gacggggtct cactctgtcg cccaggctgg 960 agtggtgtga tctcggctcgctgcagcctc cacctcccgg gcccaatcgg ttctcccgcc 1020 tcagcctcct gggtggctgggactgctggc ccgtgccacc acgcttggct aatttttttt 1080 tattattgta ttttttgtaaagatggagtt tcacctcttt gcctgggcag gtctcaaact 1140 cctgagatca aatgatcctccccccttggc ctcccaaagt gcgtggatta taggcatgag 1200 ccattgtatc tggctagcatgggagttttg aactgtccca tttccaacct gggccagtgc 1260 attcctcctt aggcagcctggtggtccctg ctcctgggat gtcactatat tgatgctgaa 1320 cttagtgcag acacctgatctgcctagcgt actgcaaccc agagctcctg ggcccaggcg 1380 atcctcctgt ctcagcctcctgagtagctg ggactctagg cacacaccac tatgcgtggc 1440 tctccatgct tcttgggtctaccctctgag atgtttttcc ttttctttca ccttccttga 1500 ttccttctga agagggcgttgcacaatgtg ctgcttttga tggttgagca aatttctcag 1560 cctccttcct gcctatagagagttggggca ggctgggcgc cagctcacgc ctgtaatccc 1620 agggaggctg aggcgggcagatcacgaggt caggacatca agaccggcct ggccgacatg 1680 gtgggacccc atctctactaacaatacaaa aattggctgg gtatggtggc acgtgcctgt 1740 ggtcccggct gctggggaggctgaggcggg agagttgctt gggcccggga ggcggaggtt 1800 gcagtggcgg gagaattgcttggggcccgg gaggcggagg ttgcggtgag ccgagattgt 1860 gccagtgcac actgcactccagcctggtga cagagtgaga ctccgtcttc aaaaa 1915 41 1809 DNA Homo sapiensmisc_feature Incyte ID No 5674114CB1 41 atgggcctgg ccagggccct acgccgcctcagcggcgccc tggattcggg agacagccgg 60 gcgggcgatg aagaggaggc cgggcccgggttgtgccgca acgggtgggc gccggcaccg 120 gtgcagtcac ccgtgggccg gcgccgcggtcgcttcgtca agaaagacgg gcactgcaac 180 gtgcgtttcg taaacctggg tggccagggcgcgcgctacc tgagcgacct gttcaccaca 240 tgcgtggacg tgcgctggcg ctggatgtgcctgctcttct cctgctcctt cctcgcctcc 300 tggctgctct tcggcctggc cttctggctcattgcctcgc tgcacggcga cctggccgcc 360 ccgccaccgc ccgcgccctg cttctcacacgtggccagct tcctggccgc cttcctcttc 420 gcgctggaga cgcagacgtc catcggctacggcgtgcgca gcgtcaccga ggagtgcccg 480 gccgctgtgg ccgccgtggt gctgcagtgcattgccggct gcgtgctcga cgccttcgtc 540 gtgggtgctg tcatggccaa gatggccaaacccaagaagc gcaacgagac gctggtcttc 600 agcgagaacg ccgtcgtggc gctgcgcgaccaccgcctct gcctcatgtg gcgcgtcggc 660 aacctgcgcc gcagccacct ggtcgaggcccacgtgcgtg cccagctgct gcagccccgt 720 gtgaccccag agggtgagta catcccgctggaccaccagg atgtggatgt gggctttgat 780 ggaggcaccg atcgtatctt cctcgtgtcccccatcacca tcgtccatga gatcgactct 840 gccagtcctc tgtatgagct aggacgtgccgagctggcca gggctgactt tgagctggtg 900 gtcattctcg aggggatggt tgaggccacagccatgacca cacagtgtcg ctcgtcctac 960 ctccctggtg aactgctctg gggccatcgttttgagccag ttctcttcca gcgtggctcc 1020 cagtatgagg tcgactatcg ccacttccatcgcacttatg aggtcccagg gacaccggtc 1080 tgcagtgcta aggagctgga tgaacgggcagagcaggctt cccacagcct caagtctagt 1140 ttccccggct ctctgactgc attttgttatgagaatgaac ttgctctgag ctgctgccag 1200 gaggaagatg aggacgatga gactgaggaagggaatgggg tggaaacaga agatggggct 1260 gctagccccc gagttctcac accaaccctggcgctgaccc tgcctccatg atgcaaactg 1320 atgtcccctt ccccgtgtat gcccccttccccaaggtagc aagatggagg gatggggctc 1380 tctcctggga tgggggcagg tgttcctgaataccgacagg cctgctgggt aaatgactag 1440 gtggtaaggt tctgccatgc ctggtgacccaccatggaca tactggacct taattcctct 1500 gcttctgtgc tccctcctga gaaccctttatgagcctgat tcctcagtct caccagaatt 1560 ctggatcacc caagaggaaa agactggcagttctagattc ctctatatgg ggagacctgg 1620 attgttgacc agggtgagaa gccaatggtatagactgcct ctggggaagc aagttggcag 1680 ttcttgaaca gcatcagata tcaagagtttgtaggtctgg attcacctaa gattcaaggg 1740 agtgttgctt ctcaactcag ccaactgagtagcaaatcat ttgttctaga ccacctaagg 1800 agggaaggt 1809 42 1730 DNA Homosapiens misc_feature Incyte ID No 1254635CB1 42 ctttggccta ttataccatggatgctaaaa atggttctaa ctgaaaaccc aaaccaagaa 60 atagcaacaa gtctagaattcttactacta caaaactcac ctggatccct aagggcacag 120 caaagaatga gctattacggcagcagctat catattatca atgcggacgc aaaataccca 180 ggctacccgc cagagcacattatagctgag aagagaagag caagaagacg attacttcac 240 aaagatggca gctgtaatgtctacttcaag cacatttttg gagaatgggg aagctatgtg 300 gttgacatct tcaccactcttgtggacacc aagtggcgcc atatgtttgt gatattttct 360 ttatcttata ttctctcgtggttgatattt ggctctgtct tttggctcat agcctttcat 420 catggcgatc tattaaatgatccagacatc acaccttgtg ttgacaacgt ccattctttc 480 acaggggcct ttttgttctccctagagacc caaaccacca taggatatgg ttatcgctgt 540 gttactgaag aatgttctgtggccgtgctc atggtgatcc tccagtccat cttaagttgc 600 atcataaata cctttatcattggagctgcc ttggccaaaa tggcaactgc tcgaaagaga 660 gcccaaacca ttcgtttcagctactttgca cttataggta tgagagatgg gaagctttgc 720 ctcatgtggc gcattggtgattttcggcca aaccacgtgg tagaaggaac agttagagcc 780 caacttctcc gctatacagaagacagtgaa gggaggatga cgatggcatt taaagacctc 840 aaattagtca acgaccaaatcatcctggtc accccggtaa ctattgtcca tgaaattgac 900 catgagagcc ctctgtatgcccttgaccgc aaagcagtag ccaaagataa ctttgagatt 960 ttggtgacat ttatctatactggtgattcc actggaacat ctcaccaatc tagaagctcc 1020 tatgttcccc gagaaattctctggggccat aggtttaatg atgtcttgga agttaagagg 1080 aagtattaca aagtgaactgcttacagttt gaaggaagtg tggaagtata tgcccccttt 1140 tgcagtgcca agcaattggactggaaagac cagcagctcc acatagaaaa agcaccacca 1200 gttcgagaat cctgcacgtcggacaccaag gcgagacgaa ggtcatttag tgcagttgcc 1260 attgtcagca gctgtgaaaaccctgaggag accaccactt ccgccacaca tgaatatagg 1320 gaaacacctt atcagaaagctctcctgact ttaaacagaa tctctgtaga atcccaaatg 1380 tagtcctaaa ttgcaattatgagggctacc actgaatcat tttatctttc agccaatcaa 1440 gtcgttgtaa acgtggcttttttgaaagtg ttatggctat gttttatgat gatgctgggt 1500 aagtagagta agttaaacttggtaaaagat aatctaaaaa ttccatagtt ctcagttatt 1560 aaaatttttc ttgttcgccaattttgtatt aagaatgcta ttaagcctaa ttgattaaaa 1620 tttatctttt ttattatcttacatgcttgt atcttcagtt ggaggtgtag tattcaaaaa 1680 cggggaatga aggcaggaaggaggctggaa taaataaaaa taaaatgatt 1730 43 1147 DNA Homo sapiensmisc_feature Incyte ID No 1670595CB1 43 gcagctgtct tttccggccc ccgtgcactctccgcccgag gcggagcccc cggctcgcgg 60 ggatcgcccc cgagcgctgc gtcctgcgggtgggtcacct aacccatttg tggcttcctc 120 tacctgtgct cagccatggc cagcgagagctcacctctgc tggcctaccg gctcctgggg 180 gaggaggggg ttgccctccc tgccaatggggccgggggtc ctggaggggc gtctgcccgg 240 aagctgtcca ccttcctggg tgtggtggtgcccactgtcc tgtccatgtt cagcatagtt 300 gtttttctga ggattgggtt cgtggtgggtcatgctgggc tactgcaggc cctggccatg 360 ctgctggttg cctacttcat cctggcactcaccgtcctct ctgtctgtgc catcgccacc 420 aatggagccg tgcagggggg cggagcctactgtatcctcc aacatcgatg gactgggatg 480 ccacagggcc cagtgggctc cgggtcctgccccagggcta cggcttggaa cctgctgtat 540 ggctccctgc tgctgggcct tgtgggtggggtctgcacct tgggagccgg cctctatgcc 600 cgggcctcat tcctcacatt cctgctggtctctggctccc tggcctctgt gctcatcagt 660 tttgtggctg tggggccgag ggacatccgcttgactccta ggcctggccc caatggctcc 720 tccctgccgc cccggtttgg ccacttcaccggcttcaaca gcagtaccct gaaggacaac 780 ttgggcgctg gctatgctga ggactacaccacgggagccg tgatgaattt tgccagcgtc 840 tttgctgtcc tctttaacgg caggcatcatggctggggcc aacatgtcag gggagctgaa 900 ggaccccagc cgggcgatcc ctctgggcacgatcgtcgcc gtcgcctaca ccttcttcgt 960 ctatgccctg cttttctttc tctccagcctcccttcactg gtgccttgat gctaggggcc 1020 aggcctcctc tgtgactctg ggctacctcagtttccccat tttggccaga ctcaccggcc 1080 caccggggtg gtgatgtttt cgttctgttttatttttcta actctgcatg accatgaata 1140 aaagacc 1147 44 2745 DNA Homosapiens misc_feature Incyte ID No 1859560CB1 44 cggcgacgcc agggaccccacgcatcccga gtgaagcaac tagaactcca gggctgtgaa 60 agccacaggt gggggctgagcgaggcgtgg cctcaggagc ggaggacccc ccactctccc 120 tcgagcgccg cagtccaccgtagcgggtgg agcccgcctt ggtgcgcagt tggaaaacct 180 cggagccccg ctggatctcctggctgccac ccgcaccccc cgccagccta cgccccaccg 240 tagagatgcc ttcttcggtgacggcgctgg gtcaggccag gtcctctggc cccgggatgg 300 ccccgagcgc ctgctgctgctcccctgcgg ccctgcagag gaggctgccc atcctggcgt 360 ggctgcccag ctactccctgcagtggctga agatggattt cgtcgccggc ctctcagttg 420 gcctcactgc cattccccaggcgctggcct atgctgaagt ggctggactc ccgccccagt 480 atggcctcta ctctgccttcatgggctgct tcgtgtattt cttcctgggc acctcccggg 540 atgtgactct gggccccaccgccattatgt ccctcctggt ctccttctac accttccatg 600 agcccgccta cgctgtgctgctggccttcc tgtccggctg catccagctg gccatggggg 660 tcctgcgttt ggggttcctgctggacttca tttcctaccc cgtcattaaa ggcttcacct 720 ctgctgctgc cgtcaccatcggctttggac agatcaagaa cctgctggga ctacagaaca 780 tccccaggcc gttcttcctgcaggtgtacc acaccttcct caggattgca gagaccaggg 840 taggtgacgc cgtcctggggctggtctgca tgctgctgct gctggtgctg aagctgatgc 900 gggaccacgt gcctcccgtccaccccgaga tgccccctgg tgtgcggctc agccgtgggc 960 tggtctgggc tgccacgacagctcgcaacg ccctggtggt ctccttcgca gccctggttg 1020 cgtactcctt cgaggtgactggataccagc ctttcatcct aacaggggag acagctgagg 1080 ggctccctcc agtccggatcccgcccttct cagtgaccac agccaacggg acgatctcct 1140 tcaccgagat ggtgcaggacatgggagccg ggctggccgt ggtgcccctg atgggcctcc 1200 tggagagcat tgcggtggccaaagccttcg catctcagaa taattaccgc atcgatgcca 1260 accaggagct gctggccatcggtctcacca acatgttggg ctccctcgtc tcctcctacc 1320 cggtcacagg cagctttggacggacagccg tgaacgctca gtcgggggtg tgcaccccgg 1380 cggggggcct ggtgacgggagtgctggtgc tgctgtctct ggactacctg acctcactgt 1440 tctactacat ccccaagtctgccctggctg ccgtcatcat catggccgtg gccccgctgt 1500 tcgacaccaa gatcttcaggacgctctggc gtgttaagag gctggacctg ctgcccctgt 1560 gcgtgacctt cctgctgtgcttctgggagg tgcagtacgg catcctggcc ggggccctgg 1620 tgtctctgct catgctcctgcactctgcag ccaggcctga gaccaaggtg tcagaggggc 1680 cggttctggt cctgcagccggccagcggcc tgtccttccc tgccatggag gctctgcggg 1740 aggagatcct aagccgggccctggaagtgt ccccgccacg ctgcctggtc ctggagtgca 1800 cccatgtctg cagcatcgactacactgtgg tgctgggact cggcgagctc ctccaggact 1860 tccagaagca gggcgtcgccctggcctttg tgggcctgca ggtccccgtt ctccgtgtcc 1920 tgctgtccgc tgacctgaaggggttccagt acttctctac cctggaagaa gcagagaagc 1980 acctgaggca ggagccagggacccagccct acaacatcag agaagactcc attctggacc 2040 aaaaggttgc cctgctcaaggcataatggg gccacccgtg ggcatccaca gtttgcaggg 2100 tgttccggaa ggttcttgtcactgtgattg gatgctggat gccgcctgat agacatgctg 2160 gcctggctga gaaacccctgagcaggtaac ccagggaaga gaaggaagcc aggcctggag 2220 gtccacggca gtgggagtggggctcactgg cttcctgtgg gatgactgga aaatgacctc 2280 gctgctgttc cctggcatgaccctctttgg aagagtggtt tggagagagc cttctagaat 2340 gacagactgt gcgaggaagcaggggcaggg gtttccagcc cgggctgtgc gaggcatcct 2400 ggggctggca gcaccttcccggctcaccag tgccacctgc gggggaggga cggggcaggc 2460 aggagtctgg gaggcgggtccgctcctctt gtctgcggca tctgtgctct ccgagagaaa 2520 accaaggtgt gtcaaatgacgtcaagtctc tatttaaaaa taattttgtg ttttctaaat 2580 ggaaaaagtg atagctttggtgattttgta aaagtcataa atgcttattg taaaaaatac 2640 aggaaaccac ccctcaccctgtccacttgg gtgatcattc cagacccctc cccaaacatg 2700 catatgtacc tgtccgtcagtgtgtggatg tatgtttaca gttct 2745 45 3204 DNA Homo sapiens misc_featureIncyte ID No 5530164CB1 45 cgacctctgg agctactgcg cctgcaagcc cagcctctctgcgccgcagg ctgcggggcc 60 agctggcgcc gcacaaatac ggggcgggac acggggcgggacacgggccg gtcccggggg 120 agggcctgag ccgcacagcc cgcccagggg tggtgcgtgtaaacgggcgt ctggatcccc 180 gaatggttgc gtgtttccgt gtgtgggtcc gggggaggcccacgaacgcc agcgaaaccg 240 ctgacaccac cgcccaacta tgaactcatc aggcgcctgaagaccgacac gccgaacatg 300 cgccgcgcgc actcgcgcac gagtgagatc atcgcgccccggtcgtgagt gcgctcacac 360 gcagcctgag actcgacggg agggggtcac gtggaagtatctgagagagg cgtacttggc 420 cactaggaaa gcacctcccc ctttccaaaa atgctccggaagtgccttcg ccctccgtaa 480 agatggccgg ggcagtcggc acgagggagg cggggatgcgcctgcgcaac aagttcggcg 540 gggaagatgg cggatgacaa ggattctctg cctaagcttaaggacctggc atttctcaag 600 aaccagctgg aaagcctgca gcggcgtgta gaagacgaagtcaacagtgg agtgggccag 660 gatggctcgc tgttgtcctc cccgttcctc aagggattcctggctggcta tgtggtggcc 720 aaactgaggg catcagcagt attgggcttt gctgtgggcacctgcactgg catctatgcg 780 gctcaggcat atgctgtgcc caacgtggag aagacattaagggactattt gcagttgcta 840 cgcaaggggc ccgactagct ctaggtgcca tggaagaggcaggatgagca gctcagcctt 900 caggtggaga cactttatct ggattcccca gctgtcatccatttgctatc tccaactttc 960 ctgccacctt catccttgcc tcccttcctg cagattgtggacagtagttc ctcagcctgc 1020 accctggatt ccttcttccc cttcctagct ccatgggactcgccccaaga ctgtggcttc 1080 aaggaccacc agccccttac tcttcaagcc ctgactgtggagttggtaga tgcctctgat 1140 cctcagtatt ctctctggca atgttccacg gcttctccttcctgggagct ggctccataa 1200 cttgattttc cccaaacgtg ttgcaatccc tgctgccccttagccaccca gggtcttgtg 1260 tgggtatgag tgtagaggat gggggtatgc caggcctgggccgtcccagg caggcccgct 1320 ggaccctgat gctactccta tccactgcca tgtacggtgcccatgcccca ttgctggcac 1380 tgtgccatgt ggacggccga gtgcccttcc ggccctcctcagccgtgctg ctgactgagc 1440 tgaccaagct actgttatgc gccttctccc ttctggtaggctggcaagca tggccccagg 1500 ggcccccacc ctggcgccag gctgctccct tcgcactatcagccctgctc tatggcgcta 1560 acaacaacct ggtgatctat cttcagcgtt acatggaccccagcacctac caggtgctga 1620 gtaatctcaa gattggaagc acagctgtgc tctactgcctctgcctccgg caccgcctct 1680 ctgtgcgtca ggggttagcg ctgctgctgc tgatggctgcgggagcctgc tatgcagcag 1740 ggggccttca agttcccggg aacacccttc ccagtccccctccagcagct gctgccagcc 1800 ccatgcccct gcatatcact ccgctaggcc tgctgctcctcattctgtac tgcctcatct 1860 caggcttgtc gtcagtgtac acagagctgc tcatgaagcgacagcggctg cccctggcac 1920 ttcagaacct cttcctctac acttttggtg tgcttctgaatctaggtctg catgctggcg 1980 gcggctctgg cccaggcctc ctggaaggtt tctcaggatgggcagcactc gtggtgctga 2040 gccaggcact aaatggactg ctcatgtctg ctgtcatgaagcatggcagc agcatcacac 2100 gcctctttgt ggtgtcctgc tcgctggtgg tcaacgccgtgctctcagca gtcctgctac 2160 ggctgcagct cacagccgcc ttcttcctgg ccacattgctcattggcctg gccatgcgcc 2220 tgtactatgg cagccgctag tccctgacaa cttccaccctgattccggac cctgtagatt 2280 gggcgccacc accagatccc cctcccaggc cttcctccctctcccatcag cagccctgta 2340 acaagtgcct tgtgagaaaa gctggagaag tgagggcagccaggttattc tctggaggtt 2400 ggtggatgaa ggggtacccc taggagatgt gaagtgtgggtttggttaag gaaatgctta 2460 ccatccccca cccccaacca agttcttcca gactaaagaattaaggtaac atcaatacct 2520 aggcctgaga aataacccca tccttgttgg gcagctccctgctttgtcct gcatgaacag 2580 agttgatgaa agtggggtgt gggcaacaag tggctttccttgcctacttt agtcacccag 2640 cagagccact ggagctggct agtccagccc agccatggtgcatgactctt ccataaggga 2700 tcctcaccct tccactttca tgcaagaagg cccagttgccacagattata caaccattac 2760 ccaaaccact ctgacagtct cctccagttc cagcaatgcctagagacatg ctccctgccc 2820 tctccacagt gctgctcccc acacctagcc tttgttctggaaaccccaga gagggctggg 2880 cttgactcat ctcagggaat gtagcccctg ggccctggcttaagccgaca ctcctgacct 2940 ctctgttcac cctgagggct gtcttgaagc ccgctacccactctgaggct cctaggaggt 3000 accatgcttc ccactctggg gcctgcccct gcctagcagtctcccagctc ccaacagcct 3060 ggggaagctc tgcacagagt gacctgagac caggtacaggaaacctgtag ctcaatcagt 3120 gtctctttaa ctgcataagc aataagatct taataaagtcttctaggctg tagggtggtt 3180 cctacaacca cagccaaaaa aaaa 3204 46 2763 DNAHomo sapiens misc_feature Incyte ID No 139115CB1 46 tgcatttgctatgactttga ccggtccact gacaacgcaa tatgtttatc ggagaatatg 60 ggaagaaactggcaactaca ctttttcatc tgatagcaat atttctgagt gtgaaaaaaa 120 caaaagcagcccaatttttg cattccagga ggaagttcag aaaaaagtgt cacgttttaa 180 tctgcagatggacataagtg gattaattcc tggtctagtg tctacattca tacttttgtc 240 tattagtgatcactacggac gaaaattccc tatgattttg tcttccgttg gtgctcttgc 300 aaccagcgtttggctctgtt tgctttgcta ttttgccttt ccattccagc ttttgattgc 360 atctaccttcattggtgcat tttgtggcaa ttataccaca ttttggggag cttgctttgc 420 ctatatagttgatcagtgta aagaacacaa acaaaaaaca attcgaatag ctatcattga 480 ctttctacttggacttgtta ctggactaac aggactgtca tctggctatt ttattagaga 540 gctaggttttgagtggtcgt ttctaattat tgctgtgtct cttgctgtta atttgatcta 600 tattttattttttctcggag atccagtgaa agagtgttca tctcagaatg ttactatgtc 660 atgtagtgaaggcttcaaaa acctatttta ccgaacttac atgcttttta agaatgcttc 720 tggtaagagacgatttttgc tctgtttgtt actttttaca gtaatcactt atttttttgt 780 ggtaattggcattgccccaa tttttatcct ttatgaattg gattcaccac tctgctggaa 840 tgaagtttttataggttatg gatcagcttt gggtagtgcc tcttttttga ctagtttcct 900 aggaatatggcttttttctt attgtatgga agatattcat atggccttca ttgggatttt 960 taccacgatgacaggaatgg ctatgaccgc gtttgccagt acaacactga tgatgttttt 1020 agccagggtgccgttccttt tcactattgt gccattctct gttctacggt ccatgttgtc 1080 aaaagtggttcgttcgactg aacaaggtac cctgtttgct tgtattgctt tcttagaaac 1140 acttggaggagtcactgcag tttctacttt taatggaatt tactcagcca ctgttgcttg 1200 gtaccctggcttcactttcc tgctgtctgc tggtctgtta ctacttccag ccatcagtct 1260 atgtgttgtcaagtgtacca gctggaatga gggaagctat gaacttctta tacaagaaga 1320 atccagtgaagatgcttcag acaggtgact gtgatttaaa caaacaaaaa aaatctatga 1380 atgcacatatcatataccat gacttctgaa gactataaat gaattccaca atcagtgctt 1440 cactgagaaccaattttacc tatcttttct tctaaactga acagtcagag agacagctcc 1500 tggctttagcttcttgtggt accacgcact ttgagcactt tgtgcgtatc atgcaatata 1560 cttgcaatacacagaacaaa tttcaaatac gcctcacttt tagacttaga agagaaacat 1620 taaaacttaagggtgtaagg agggatcaag aaacttgata aggtcaaaag caataatctc 1680 tctgacatattccaggctct tacactgaga ccaaagagaa atctttacct cagtttcttc 1740 atcagcagaatgggtttctg gcctctctca gggataattt tgaaggcata atgaaaatta 1800 tgatgaatcactcattggta ggaaaataat gatataagtt tcaaatatgt atgattttac 1860 ctatacttggtaatgctttg ttttatagag cctgttaagc tgctattgat agtcggagct 1920 tatatactgtgacttctgaa gactatacat gaattccaca atcagtgctt tgttgataca 1980 aaatccttaaaagggaggca ctttaaagaa tatgtatttt tcacttttct taatatgttt 2040 catcggtgacaggcatgata atatttctat atgtaatggg taattgggaa aaaatagatg 2100 ataaataaaattgctctaaa gaagttaaaa aactgaatga acagctaata ctggtataaa 2160 gtaactaatgtttggagcca acatttgttc cttgtgtcag caaaaggata ttcacattcc 2220 atgatccctggctgagaatt ctgcctctag tctttcttac ccagctgttg tctatccttg 2280 ttcaattataaatactgcta agggcatttt taaaatacga tcttgtagtc cttaaatttg 2340 aatccgtcagcacggtcact cataggaaaa tgatcaaaca agcaagccag tcatgatttg 2400 actccttcccatctcatttc ttactgcctt acgctcatcc tgaggtccac cttggtctct 2460 aaaaacaccatgtgttctca tgcctccatg tcttttcaca cactgttcca tttgctcttc 2520 ctcccacattacattgaaac tttcaagcct cagtcgaaac attgcttctt ctggatagca 2580 gccttcttgacatccctcct cactccccag tccctacagg gcttccatag ctctttgtgt 2640 gcacttcgatcccagcattt tccatcgact tgtaattgtt tctgctacct gacaatcatc 2700 gccttgagtactgggacaac ctttgattac tcattatatc ctcaataaat atttgttgaa 2760 cta 2763 471639 DNA Homo sapiens misc_feature Incyte ID No 1702940CB1 47 atcgcactgaggcttgagtc tgacttctct cccccacctg ctgtgccctt aaactgcaga 60 gatcggggcgggggttgggg ggcaagcggc tcagatgggt tcaaaaaact ccccaggctc 120 aactctggttctgactgcct gagacatggg cagctgacac agcagacctt gaatcctgag 180 gatgtgaggcagggtatatc tgggaggccg gaggacgtgt ctggttatta cacagatgca 240 cagctggacgtgggatccac acagctcaga acagttggat cttgctcagt ctctgtcaga 300 ggaagatcccttggacaaga ggaccctgcc ttggtgtgag agtgagggta gaggaagctg 360 gaacgagggttaaggaaaac cttccagtct ggacagtgac tggagagctc caaggaaagc 420 ccctcggtaacccagccgct ggcaccatga acccagagag cagtatcttt attgaggatt 480 accttaagtatttccaggac caagtgagca gagagaatct gctacaactg ctgactgatg 540 atgaagcctggaatggattc gtggctgctg ctgaactgcc cagggatgag gcagatgagc 600 tccgtaaagctctgaacaag cttgcaagtc acatggtcat gaaggacaaa aaccgccacg 660 ataaagaccagcagcacagg cagtggtttt tgaaagagtt tcctcggttg aaaagggagc 720 ttgaggatcacataaggaag ctccgtgccc ttgcagagga ggttgagcag gtccacagag 780 gcaccaccattgccaatgtg gtgtccaact ctgttggcac tacctctggc atcctgaccc 840 tcctcggcctgggtctggca cccttcacag aaggaatcag ttttgtgctc ttggacactg 900 gcatgggtctgggagcagca gctgctgtgg ctgggattac ctgcagtgtg gtagaactag 960 taaacaaattgcgggcacga gcccaagccc gcaacttgga ccaaagcggc accaatgtag 1020 caaaggtgatgaaggagttt gtgggtggga acacacccaa tgttcttacc ttagttgaca 1080 attggtaccaagtcacacaa gggattggga ggaacatccg tgccatcaga cgagccagag 1140 ccaaccctcagttaggagcg tatgccccac ccccgcatgt cattgggcga atctcagctg 1200 aaggcggtgaacaggttgag agggttgttg aaggccccgc ccaggcaatg agcagaggaa 1260 ccatgatcgtgggtgcagcc actggaggca tcttgcttct gctggatgtg gtcagccttg 1320 catatgagtcaaagcacttg cttgaggggg caaagtcaga gtcagctgag gagctgaaga 1380 agcgggctcaggagctggag gggaagctca actttctcac caagatccat gagatgctgc 1440 agccaggccaagaccaatga ccccagagca gtgcagccac cagggcagaa atgccgggca 1500 caggccaggacaaaatgcag actttttttt ttttcaagtc tttgacgggg aagggagctc 1560 cgctttttcccccagtaggg gtggcggggc ccaactctgg gccgtgtgaa cctcccgggg 1620 ggggggattcgattaacgc 1639 48 1600 DNA Homo sapiens misc_feature Incyte ID No1703342CB1 48 caaggcggcc caggacaggc aggggctgca cgcggtgaag aaaccaagacgcagagaggc 60 caagcccctt gccttgggtc acacagccaa aggaggcaga gccagaactcacaaccagat 120 ccagaggcaa cagggacatg gccacctggg acgaaaaggc agtcacccgcagggccaagg 180 tggctcccgc tgagaggatg agcaagttct taaggcactt cacggtcgtgggagacgact 240 accatgcctg gaacatcaac tacaagaaat gggagaatga agaggaggaggaggaggagg 300 agcagccacc acccacacca gtctcaggcg aggaaggcag agctgcagcccctgacgttg 360 cccctgcccc tggccccgca cccagggccc cccttgactt caggggcatgttgaggaaac 420 tgttcagctc ccacaggttt caggtcatca tcatctgctt ggtggttctggatgccctcc 480 tggtgcttgc tgagctcatc ctggacctga agatcatcca gcccgacaagaataactatg 540 ctgccatggt attccactac atgagcatca ccatcttggt cttttttatgatggagatca 600 tctttaaatt atttgtcttc cgcctggagt tctttcacca caagtttgagatcctggatg 660 ccgtcgtggt ggtggtctca ttcatcctcg acattgtcct cctgttccaggagcaccagt 720 ttgaggctct gggcctgctg attctgctcc ggctgtggcg ggtggcccggatcatcaatg 780 ggattatcat ctcagttaag acacgttcag aacggcaact cttaaggttaaaacagatga 840 atgtacaatt ggccgccaag attcaacacc ttgagttcag ctgctctgagaaggaacaag 900 aaattgaaag acttaacaaa ctattgcgac agcatggact tcttggtgaagtgaactaga 960 cccggaccag ctcccctcaa aaagaagaca ctgtctcatg ggcctgtgctgtcacgagag 1020 gaacagctgc ccctcctggg ccgcttggtg agaggtttgg tttgatacctctgcctccct 1080 cctgccagca tggattctgg gtggacacag ccttgtggaa ggtccagtaccaccaagagc 1140 tgcccatcca ctcccacccc acactgtatc aaatgtatca cattttctcatgttgaacac 1200 tttagcctta attgaaaatg agcaacaaag ctggacaatt gctagttgtatataaaattt 1260 aatctcaccg aatgtacagt tttcaaattt cacgtgtata ttaaggaactgatgcatctg 1320 agcattctga aagaaagaaa aagaagctac tttagctgcc accccattctagaaaagtct 1380 cttattttca agctgttcta aatagcttcg tctcagtttc cccaaaaggggtacccaggc 1440 ccctcctctg tgtgccccag ctgcatcagc cagcttctag gtggctccattgttttctgc 1500 cacctgacaa catttttcct caattactgt acaactactg tataaaataaaacaactact 1560 gtataaaata aactctctct tttccctgga aaaaaaaaaa 1600 49 2380DNA Homo sapiens misc_feature Incyte ID No 1727529CB1 49 ctgagccatggggggaaagc agcgggacga ggatgacgag gcctacggga agccagtcaa 60 atacgacccctcctttcgag gccccatcaa gaacagaagc tgcacagatg tcatctgctg 120 cgtcctcttcctgctcttca ttctaggtta catcgtggtg gggattgtgg cctggttgta 180 tggagacccccggcaagtcc tctaccccag gaactctact ggggcctact gtggcatggg 240 ggagaacaaagataagccgt atctcctgta cttcaacatc ttcagctgca tcctgtccag 300 caacatcatctcagttgctg agaacggcct acagtgcccc acaccccagg tgtgtgtgtc 360 ctcctgcccggaggacccat ggactgtggg aaaaaacgag ttctcacaga ctgttgggga 420 agtcttctatacaaaaaaca ggaacttttg tctgccaggg gtaccctgga atatgacggt 480 gatcacaagcctgcaacagg aactctgccc cagtttcctc ctcccctctg ctccagctct 540 gggacgctgctttccatgga ccaacattac tccaccggcg ctcccaggga tcaccaatga 600 caccaccatacagcagggga tcagcggtct tattgacagc ctcaatgccc gagacatcag 660 tgttaagatctttgaagatt ttgcccagtc ctggtattgg attcttgttg ccctgggggt 720 ggctctggtcttgagcctac tgtttatctt gcttctgcgc ctggtggctg ggcccctggt 780 gctggtgctgatcctgggag tgctgggcgt gctggcatac ggcatctact actgctggga 840 ggagtaccgagtgctgcggg acaagggcgc ctccatctcc cagctgggtt tcaccaccaa 900 cctcagtgcctaccagagcg tgcaggagac ctggctggcc gccctgatcg tgttggcggt 960 gcttgaagccatcctgctgc tggtgctcat cttcctgcgg cagcggattc gtattgccat 1020 cgccctcctgaaggaggcca gcaaggctgt gggacagatg atgtctacca tgttctaccc 1080 actggtcacctttgtcctcc tcctcatctg cattgcctac tgggccatga ctgctctgta 1140 cctggctacatcggggcaac cccagtatgt gctctgggca tccaacatca gctcccccgg 1200 ctgtgagaaagtgccaataa atacatcatg caaccccacg gcccaccttg tgaactcctc 1260 gtgcccagggctgatgtgcg tcttccaggg ctactcatcc aaaggcctaa tccaacgttc 1320 tgtcttcaatctgcaaatct atggggtcct ggggctcttc tggaccctta actgggtact 1380 ggccctgggccaatgcgtcc tcgctggagc ctttgcctcc ttctactggg ccttccacaa 1440 gccccaggacatccctacct tccccttaat ctctgccttc atccgcacac tccgttacca 1500 cactgggtcattggcatttg gagccctcat cctgaccctt gtgcagatag cccgggtcat 1560 cttggagtatattgaccaca agctcagagg agtgcagaac cctgtagccc gctgcatcat 1620 gtgctgtttcaagtgctgcc tctggtgtct ggaaaaattt atcaagttcc taaaccgcaa 1680 tgcatacatcatgatcgcca tctacgggaa gaatttctgt gtctcagcca aaaatgcgtt 1740 catgctactcatgcgaaaca ttgtcagggt ggtcgtcctg gacaaagtca cagacctgct 1800 gctgttctttgggaagctgc tggtggtcgg aggcgtgggg gtcctgtcct tctttttttt 1860 ctccggtcgcatcccggggc tgggtaaaga ctttaagagc ccccacctca actattactg 1920 gctgcccatcatgacctcca tcctgggggc ctatgtcatc gccagcggct tcttcagcgt 1980 tttcggcatgtgtgtggaca cgctcttcct ctgcttcctg gaagacctgg agcggaacaa 2040 cggctccctggaccggccct actacatgtc caagagcctt ctaaagattc tgggcaagaa 2100 gaacgaggcgcccccggaca acaagaagag gaagaagtga cagctccggc cctgatccag 2160 gactgcaccccacccccacc gtccagccat ccaacctcac ttcgccttac aggtctccat 2220 tttgtggtaaaaaaaggttt taggccaggc gccgtggctc acgcctgtaa tccaacactt 2280 tgagaggctgaggcgggcgg atcacctgag tcaggagttc gagaccagcc tggccaacat 2340 ggtgaaacctccgtctctat taaaaataca aaaattagcc 2380 50 3038 DNA Homo sapiensmisc_feature Incyte ID No 2289333CB1 50 aggggcaggg aggcgggcac caggcgcgggtccctccggg caggcgaggt aggcctgggc 60 ctgacgccgg ccacgcagcg gcgggagagtgagcactcgg gcggcggcgt cctggagacc 120 cgcgagagat ggaagcggcg gcgacgccggcggctgccgg ggcggcgagg cgcgaggagc 180 tagatatgga tgtaatgagg cccttgataaatgagcagaa ttttgatggg acatcagatg 240 aagaacatga gcaagagctt ctgcctgttcagaagcatta ccaacttgat gatcaagagg 300 gcatttcatt tgtacaaact cttatgcaccttcttaaagg aaatattgga actggccttt 360 taggacttcc attggcaata aaaaatgcaggcatagtgct tggaccaatc agccttgtgt 420 ttataggaat tatttctgtt cactgtatgcacatattggt acgttgcagt cactttctat 480 gtctgaggtt taaaaagtca acattaggttatagtgacac tgtgagcttt gctatggaag 540 tgagtccttg gagttgtctt cagaagcaagcagcatgggg gcggagtgtg gttgactttt 600 ttctggtgat aacacagctg ggattctgtagtgtttatat tgtcttctta gctgaaaatg 660 tgaaacaagt tcatgaagga ttcctggagagtaaagtgtt tatttcaaat agtaccaatt 720 catcaaaccc ttgtgagaga agaagtgttgacctaaggat atatatgctt tgctttcttc 780 catttataat tcttttggtc ttcattcgtgaactaaagaa tctatttgta ctttcattcc 840 ttgccaacgt ttccatggct gtcagtcttgtgataattta ccagtatgtt gtcaggaaca 900 tgccagatcc ccacaacctt ccaatagtggctggttggaa gaaataccca ctcttttttg 960 gtactgctgt atttgctttt gaaggcataggagtggtcct tccactggaa aaccaaatga 1020 aagaatcaaa gcgtttccct caagcgttgaatattggcat ggggattgtt acaactttgt 1080 atgtaacatt agctacttta ggatatatgtgtttccatga tgaaatcaaa ggcagcataa 1140 ctttaaatct tccccaagat gtatggttatatcaatcagt gaaaattcta tattcctttg 1200 gcatttttgt gacatattca attcagttctatgttccagc agagatcatt atccctggga 1260 tcacatccaa atttcatact aaatggaagcaaatctgtga atttgggata agatccttct 1320 tggttagtat tacttgtgcc ggagcaattcttattcctcg tttagacatt gtgatttcct 1380 tcgttggagc tgtgagcagc agcacattggccctaatcct gccacctttg gttgaaattc 1440 ttacattttc gaaggaacat tataatatatggatggtcct gaaaaatatt tctatagcat 1500 tcactggagt tgttggcttc ttattaggtacatatataac tgttgaagaa attatttatc 1560 ctactcccaa agttgtagct ggcactccacagagtccttt tctaaatttg aattcaacat 1620 gcttaacatc tggtttgaaa tagtaaaagcagaatcatga gtcttctatt tttgtcccat 1680 ttctgaaaat tatcaagata actagtaaaatacattgcta tatacataaa aatggtaaca 1740 aactctgttt tctttggcac gatattaatattttggaagt aatcataact ctttaccagt 1800 agtggtaaac ctatgaaaaa tccttgcttttaagtgttag caatagttca aaaaattaag 1860 ttctgaaaat tgaaaaaatt aaaatgtaaaaaaattaaag aataaaaata cttctattat 1920 tcttttatct cagtaagaaa taccttaaccaagatatctc tcttttatgc tactcttttg 1980 ccactcactt gagaacagaa taggatttcaacaataagag aataaaataa gaacatgtat 2040 aacaaaaagc tctctccaga tcatccctgtgaatgccaaa gtaaacttta tgtacagtgt 2100 aaaaaaaaaa aatctcagtt atgtttttattagccaaatt ctaatgattg gctcctggaa 2160 gtatagaaaa ctcccattaa cataatataagcatcagaaa attgcaaaca ctagaattaa 2220 ttttacactc taatggtagt tgatcttcatagtcaagagg cactgttcaa gatcatgact 2280 tagtgtttca atgaaatttg acaagggactttaaaactta tccagtgcaa ctcccttgtt 2340 tttcgtcaga ggaaaaggag gcctagaaaggttaagtaac ttggtcgaga ccactcagcc 2400 ttgagatcaa gaaaacctaa tcttctgactcccaggccag gatgttttat ttctcacatc 2460 atgtccaaga aaaagaataa attatgttcagcttaatttt agtgttgaat ctatttgatt 2520 atattttaat actttgaaaa tgaatgtgtgatttttaata gtatatgtga cctgagcaga 2580 aaatcaggga actccaagaa gcctacactgtggccatata aacctcagca agagaaagaa 2640 gctatgttct tttaaaacag aatagagaccgcttgctggt gaaactcctg gctagtaaga 2700 tgtgtgtcta gctatactat ttgtggcttgagctttttta attattacct tcctttcctg 2760 agttttgtag gcaccacatt cctgaatggcagaaaataga cacctcagaa aacggaggat 2820 ttgtggactc tttccagccc tgtggcttttcttatcacag ccttttattt attatgagca 2880 gaataaaaga atcagctagg tgtggtggtctgtgcttata atcccagcta ctctggagga 2940 taagttggga ggatcacttg agaggccaggagcttgagac cagcctgggc agcatagtga 3000 gacctcgact ctataaaaca taaaaaaaaaaaaaaaaa 3038 51 2608 DNA Homo sapiens misc_feature Incyte ID No2720354CB1 51 taggctaatt ttttttacag acacgatttc gccacgttgg ccaggctggtcttgaactcc 60 tgacctcaag tgatccaccc acctcagcct cccaaagtgt tgggattacaggcgtgagcc 120 actgcacctg gccaggctca tcactttttg cgcctattgc ctcgaagccagtctctgatg 180 ggacattagg gcaggggccc ttcagcctag tctgggacat gggccgctcactcagcagta 240 tgacaagcat cacctggaga acgggccagt ctcaggaggt cgttcatgccccactggcag 300 tgcactgtgc agccatagtg taaacaagag gcttaacctg aactggtctgagatcttggg 360 gaccccctac cctgtctcca gcagcctgtc cctttagctg tttgcctactggcaccccat 420 cctgagaagg catagatacc cggcccaccc tgccctggaa ttacaaaagtcttagactgt 480 gcctgagtgc ccggcctcct tgggagaccc tcctaggcag cctaagcaccagacccggga 540 gctgggtgct ctggtcctgc ctgcctgcct ctcactggac cctctccttccaggtacggc 600 ttcaggtcca gagcgtggag aagcctcagt accgcgggac gttgcactgcttcaagtcca 660 tcatcaagca agagagcgtg ctgggcctgt acaagggcct gggctcgccgctcatggggc 720 tcaccttcat caacgcgctg gtgttcgggg tgcagggcaa caccctccgggccctgggcc 780 acgactcgcc cctcaaccag ttcctggcag gtgcggcggc gggcgccatccagtgcgtca 840 tctgctgccc catggagctg gccaagacgc ggctgcagct gcaggacgcgggcccagcgc 900 gcacctacaa gggctcgctg gactgcctcg cgcagatcta cgggcacgagggtctgcgtg 960 gcgtcaaccg gggcatggtg tccacgttgc tgcgtgagac tcccagcttcggcgtctact 1020 tcctcaccta tgacgctctc acgcgggcgc tgggctgcga gccgggcgaccgcctgctgg 1080 tgcccaagct gctgttggcg ggcggtacgt caggcatcgt gtcctggctctctacctatc 1140 ctgtggacgt ggtcaagtcg cggctgcagg cggacggact gcggggcgccccgcgctacc 1200 gcggcatcct ggactgcgtg caccagagct accgcgccga gggctggcgcgtcttcacac 1260 gggggctggc gtccacgctg ctgcgcgcct tccccgtcaa cgctgccaccttcgccaccg 1320 tcacggtggt gctcacctac gcgcgcggcg aggaggccgg gcccgagggcgaggctgtgc 1380 ccgccgcccc tgcggggcct gccctggcgc agccctccag cctgtgacgctcaccccgcc 1440 ctccttcccc agggctcctt ctcagaaacc tgggacataa attggcccctgagtcgattg 1500 ccctgcttcc tgctgggatg ctgcgagctg tggagtctat cagatgtgggctgaattttg 1560 ctgatcagct gggtagtttt ggccgagaac tgcacttgcc tcagtgttctcatctatgaa 1620 ataaggaccc tcatgcccac actgtagagt cacgaagctc agagattattcccagcagca 1680 gccagcacct ggcctggctg aggccattgc accgttatcc tggaaactgaggcagacact 1740 ccagcccctt tctgggatcc tggccacgtc attgtgctcc tgccctgcaggctggctccc 1800 gggggtctct gatggccaac caaggggcca cccagggacc tctaactccacacatcctcc 1860 acccgggggg gtggtgggcc acccctctgg tctgtgttag ggacagaggaaaacttggtg 1920 tgcctcctgg tgtcacagaa ctggatcctc tgcatacccc agcttctccacatgccactg 1980 ctaggggtac cccagctgct gccactcctg ctggagggtg aactggggaccctgcaccct 2040 ccgggaagcc atggagtctg ctggaggcac catatcagcc tgcgggactagggtggggag 2100 caaacaggcc agcggtggag gtctggacag ttcaagtgtg atgcagctgtggcaaggaga 2160 aatccttccg cctctgggcc tcaggctgcc tgtccataaa atggggacatggccagctga 2220 cggacaactg agtctccggc ccacctacca ccgccagcca ggatcccccaaagtgtgcag 2280 agggctcagc agagaacagt atgggacccc ctcaccaggc ctggaacacctccagccaca 2340 aagaagccaa aggtcagtcc ctctgctccc cagcaaacgg tgcctcccaggcattctcag 2400 tgccagggct tcatccctgt gaaggcacag ggcctgctag tgggcacaggggtggctagt 2460 tggggcctgg ggcagaggag ggctgcacca ggcgtcctgg ggaatgtgctcagtgaagac 2520 gacactgggc tttgcacagc ctggtgtcgc tgtacagaaa ctgtcaagggaataaagtgt 2580 tctttgtttt ttaaaaaaaa aaaaaaaa 2608 52 3804 DNA Homosapiens misc_feature Incyte ID No 3038193CB1 52 ccctttcctg tcactggctactaccactcc caaccctcct caaagccgcc ggagcaaccc 60 ccaggtcttt actttacaatcggcaatttg acttgctctg ctgcatgtct ggagggacca 120 aggaaagtgt ggagacgctccaaggattag gtgatcggag cttgaaaaga aaaaaagcca 180 aacaaataaa caaaacccacccaccctaac aaatatgagg ctgctggaga gaatgaggaa 240 agactggttc atggtcggaatagtgctggc gatcgctgga gctaaactgg agccgtccat 300 aggggtgaat gggggaccactgaagccaga aataactgta tcctacattg ctgttgcaac 360 aatattcttt aacagtggactatcattgaa aacagaggag ctgaccagtg ctttggtgca 420 tctaaaactg catctttttattcagatctt tactcttgca ttcttcccag caacaatatg 480 gctttttctt cagcttttatcaatcacacc catcaacgaa tggcttttaa aaggtttgca 540 gacagtaggt tgcatgcctccgcctgtgtc ttctgcagtg attttaacca aggcagttgg 600 tggaaatgag ggcatcgttataacacccct gctcctgctg ctttttcttg gttcatcttc 660 ttctgtgcct ttcacatctattttttctca gctttttatg actgttgtgg ttcctctcat 720 cattggacag attgtccgaagatacatcaa ggattggctt gagagaaaga agcctccttt 780 tggtgctatc agcagcagtgtactcctcat gatcatctac acaacattct gtgacacgtt 840 ctctaaccca aatattgacctggataaatt cagccttgtt ctcatactgt tcataatatt 900 ttctatccag ctgagttttatgcttttaac tttcatcttt tcaacaagga ataattcggg 960 tttcacacca gcagacacagtggctatcat tttctgttct acacacaaat cccttacatt 1020 gggaattccg atgctgaagatcgtgtttgc aggctatgag catctctctt taatatctgt 1080 acccttgctc atctaccacccagctcagat ccttctggga agtgtgttgg tgccaacaat 1140 caagtcttgg atggtatcaaggcagaagaa actactccaa accagggggc cactggctaa 1200 cttgaataat ccagaaggcttggaatatct atccatcaaa tttgggcatt aaaataaata 1260 ccaagagtcc atcctccagggagtgaagct gacaaggccg acagtataac aaaggaggtg 1320 gactttctgt agcaatgtatatatgtacag gattgtacat actagcaatt ctgaagactt 1380 gtacttgtga atgttgcctcaatgcatatt ttattttttt acacaaaaat atgagatcct 1440 gtttaagtgc cttaaaatgtatttgacaag agcgttattt ccacaatatg ctttgttgat 1500 tactgccagg ggtggtacaatatttggggg ttaattttgc tttcctaatg caggaatcag 1560 tcatggtaag tgacaaaaagcaaacatgct ttccctgcag cacctttgtg taatacaacc 1620 ctatagtagt tactgtaatgtttgaaatga ggtcacacca tcaggaaaat gcccttctga 1680 tgacagtgaa aatttccaaagtcttattca tgcatacttt gatttactgt gtgattcttt 1740 ttttctacga ctgtgacatgcctcttcctt atcaactcag caggggtcat agatcgaata 1800 gatgctgaaa agcgtaagatatatgcattc cttgacatca tttttaaaga cattccttca 1860 aatagtttcc acacagaaattcctcactcc cattatgaga gattgtggtt atatgtctta 1920 aatttattat aagctgcttcaaagaaaggg tctgaatgtt tgaattatga gtgaaatcat 1980 gtgaaatttt gagttaaactctgtgatttg attttcaggg tctttaaaat atatcttaat 2040 atcttcttcc tctttattcaataatttctg tcttgcactt acacactcat aacagccaaa 2100 tatgaggcac aaaaatgttacaatcagttt gaaagcagca tcaattaatg gtagattcta 2160 ttcacattcc acaacccagaccaaattttt ttcctattac gcagatgtgc tgagcacttt 2220 ccagattgcc cctgttggccaaaagcagcc tgttacatcc tggaattaag cacacttaag 2280 gtatttgaga caatttattaatgaaaattt ccttggcaga tttgacaaat gttggcaata 2340 tttttttaaa agttaaatcatattgctttc atgaataaat gaaaatataa aggtcatgga 2400 tgcaaacaaa tgttacatatacacattctg tctctccaga tgaaaagaac atgcaaaacc 2460 atttaataac caaaatatcaagtaaaatta gttcccaacg gggcagcagc tttcaaatga 2520 gtgtccaata tttgcttctgctatagctgc aagaactgta actggaccca agtagagaat 2580 gaagccacgt atagaactacgagaacactt ttctgtgttt cccccatgcc gtcctgtcac 2640 atcctcttac acgtcctctcttgatttgat agacaatatt ggcatcctgg gtctcactga 2700 ggccgtgcta tgtcctcagcagctgttttt gttgtttcgt tattatgccc acaacaaaaa 2760 atcattcctt agaaactcaccaagtttatc tactgtgtaa atttatatta ttgttactac 2820 caggtctcat cttttgtcaatgtcattgaa taaatttcat aagagttatt ctcagtgtga 2880 attttaaggc taatgccagatcctgcaaaa atctatgcta accaggctgt agtacacact 2940 gttataaaga attttacttgtgtctaaaac tacagtaatt ttgcttaggt aattgtgctt 3000 acctatggag cacaggaaggctcttaggtt ttgttcctac aagtttcttt gaattttgga 3060 gtaaatggaa gtgtctgtctgtctgtcatc tatctgccct atcataaaaa tctttctccc 3120 taacattaaa atactgatccccgcccccaa cttatctacc tctattgtct aacacctata 3180 gtaggtgtga tcatgggataaaattcaact gaaaatgcta tgataacatt ttatcgtttg 3240 ctttaaaaat gtgctttgttttcaaataat ctttacatag tgaactttgg tggcgttagt 3300 gatatgttta tgcctatttcttttttttac acaaattcct tggcatattt tttcataaag 3360 aacaaaaaat aaaatcaaaatttattttta attcatgctt attgggattt aattattcag 3420 agcttaaaat attttgttatgtttatacac tgtaaagcta tctgttttat gcatttgttt 3480 tgtctaaatg tatttatgaaagaaatacat tagattatat ttatgtttac tcatttttcc 3540 acctggattt tttttaatggttgttacaaa attagatttt ttaatgggta ataatgttgg 3600 tattttcatg ttttttcttagtattaaaat ttttgtgggt tttttaaaat ttttccctat 3660 tctgttaaaa attaacacacctctagctaa tgttcagtgt ttgtgctaaa taccaaattt 3720 tttcaaaagg attggttaagtcataaagtg gattatttat gatgactgga agatgaaaat 3780 aattatatga ttaaacaaagaatg 3804 53 1894 DNA Homo sapiens misc_feature Incyte ID No 3460979CB153 acggatcact agtatgcggc gcagtgtgct ggaaagggaa caaacatggc cgctctggcg 60cccgtcggct cccccgcctc ccgcggtcct aggctggccg cgggcctccg gctgctccca 120atgctgggtt tgctgcagtt gctggccgag cctggcctgg gccgcgtcca tcacctggca 180ctcaaggatg atgtgaggca taaagttcat ctgaacacct ttggcttctt caaggatggg 240tacatggtgg tgaatgtcag tagcctctca ctgaatgagc ctgaagacaa ggatgtgact 300attggattta gcctagaccg tacaaagaat gatggctttt cttcttacct ggatgaagat 360gtgaattact gtattttaaa gaaacagtct gtctctgtca cccttttaat cctagacatc 420tccagaagtg aggtaagagt aaagtctcca ccagaagctg gtacccagtt accaaagatc 480atcttcagca gggatgagaa agtccttggt cagagccagg agcctaatgt taaccctgct 540tcagcaggca accagaccca gaagacacaa gatggtggaa agtctaaaag aagtacagtg 600gattcaaagg ccatgggaga gaaatccttt tctgttcata ataatggtgg ggcagtgtca 660tttcagtttt tctttaacat cagcactgat gaccaagaag gcctttacag tctttatttt 720cataaatgcc ttggaaaaga attgccaagt gacaagttta cattcagcct tgatattgag 780atcacagaga agaatcctga cagctacctc tcagcaggag aaattcctct ccccaaatta 840tacatctcaa tggccttttt cttctttctt tctgggacca tctggattca tatccttcga 900aaacgacgga atgatgtatt taaaatccac tggctgatgg cggcccttcc tttcaccaag 960tctctttcct tggtgttcca tgcaattgac taccactaca tctcctccca gggcttccct 1020atcgaaggct gggctgttgt gtactacata actcaccttt tgaaaggggc gctactcttc 1080atcaccattg cactcattgg cactggctgg gctttcatta agcacatcct ttctgataaa 1140gacaaaaaga tcttcatgat tgtcattcca ctccaggtcc tggcaaatgt agcctacatc 1200atcatagagt ccaccgagga gggcacgact gaatatggct tgtggaagga ctctctattt 1260ctggtcgacc tgttgtgttg tggtgccatc ctcttcccag tggtgtggtc aatcagacat 1320ttacaagaag catcagcaac agatggaaaa gctgctatta acttagcaaa gctgaaactt 1380ttcagacatt attacgtctt gattgtgtgt tacatatact tcactaggat cattgcattt 1440ctcctcaaac tcgctgttcc attccagtgg aagtggctct accagctcct ggatgaaacg 1500gccacactgg tcttctttgt tctaacgggg tataaattcc gtccggcttc agataacccc 1560tacctacaac tttctcagga agaagaagac ttggaaatgg agtccgtgta agaaatcttt 1620cttccctctt ccttagccct gaaccctttg nctaacacaa agcagcacag tgtgaatcga 1680gccggctggt ctcagcattt cgtggctgca ggggtgggtc ctctatattt agcagaaggg 1740accggcactg gagcccaagg ggtcggtctg gttgaaggca agatttggca accatactgg 1800gctgtgccgg aaaaggaaag ggggggccaa aaaacaattg gggccggcgt caaaaaaccg 1860ggcgaacaag agaaaaagcg ggcccaggag aaag 1894 54 1668 DNA Homo sapiensmisc_feature Incyte ID No 7472200CB1 54 atgacactgg tttactttcc tccttcaaagcttcagcagc agcagcagcc atcgagatcc 60 agtcgcctgg cccaacagtt ggcccaatcctcctggcagc tggccctgcg ctttggcaaa 120 cggaccacta tccacggcct ggacaggctgcttagtgcca aggccagtcg atgggagcga 180 ttcgtctggc tgtgcacctt tgtgagtgccttcctgggcg cggtgtacgt ttgcctgatt 240 ctctccgccc gctacaacgc cgcccacttccagacggtgg tggatagcac gcggtttccg 300 gtttaccgca taccatttcc ggtcataacgatctgcaacc ggaatcgcct caactggcaa 360 cgcctggcgg aggcgaagtc aagattcctggccaacggca gcaactccgc ccagcaggag 420 ctcttcgagc tgattgtggg cacctacgacgatgcttact tcggtcactt tcagtccttc 480 gagcgattgc gcaaccagcc aacggagctgctcaactatg tcaatttcag ccaggtggtg 540 gattttatga cctggcgctg caacgagctgctcgcggaat gcctgtggcg ccaccatgcc 600 tacgactgct gcgagatccg ctcgaagcggcgcagcaaga acggcttgtg ctgggctttc 660 aactcgctgg agacggaaga gggcaggcggatgcagctgc tcgatcccat gtggccctgg 720 cgtactgggt cggcgggtcc catgagcgccctctccgtgc gtgttctcat ccagcccgcg 780 aagcactggc cggggcacag ggagacgaatgccatgaagg gcatcgatgt catggttacc 840 gagccatttg tgtggcacaa caatccgttcttcgtggccg cgaacacgga gacgaccatg 900 gagatcgaac ccgtcatcta cttctatgacaacgacaccc ggggagttcg ctccgaccag 960 cgccagtgcg tcttcgatga tgagcacaacagcaaggatt tcaagtcgct gcaaggatac 1020 gtttacatga ttgaaaactg tcagtccgagtgccatcagg agtacttggt gcgctattgc 1080 aactgcacaa tggacctact gtttccaccggacctgctca tctactccca caatcccggc 1140 gagaaggagt tcgttcgcaa ccaatttcagggaatgtcct gcaagtgctt ccgcaactgc 1200 tactccctca actacatcag cgatgtccggcccgccttcc tgccaccgga tgtgtacgca 1260 aacaactcct atgtggacct ggatgtgcactttcgcttcg agaccattat ggtctatcgc 1320 accagcctcg tcttcggctg ggtggacttaatggttagct ttggaggaat tgccggtctt 1380 tttcttggct gctccctaat tagtggcatggaactggcct atttcctgtg cattgaggtg 1440 ccggcctttg ggctggatgg actgcgtcgaaggtggaagg ctcgacggca gatggatctg 1500 ggcgtaaccg tgcccacgcc cactttgaactttcaacaaa ccacgcccag tcagctgatg 1560 gagaactaca ttatgcaact gaaggctgagaaggcgcaac agcagaaggc gaactttcaa 1620 aactggcacc gcataacatt tgctcaaaagcatgttattg gcaagtga 1668

What is claimed is:
 1. An isolated polypeptide comprising an amino acidsequence selected from the group consisting of: a) an amino acidsequence selected from the group consisting of SEQ ID NO:1-27, b) anaturally occurring amino acid sequence having at least 90% sequenceidentity to an amino acid sequence selected from the group consisting ofSEQ ID NO: 1-27, c) a biologically active fragment of an amino acidsequence selected from the group consisting of SEQ ID NO:1-27, and d) animmunogenic fragment of an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27.
 2. An isolated polypeptide of claim 1selected from the group consisting of SEQ ID NO: 1-27.
 3. An isolatedpolynucleotide encoding a polypeptide of claim
 1. 4. An isolatedpolynucleotide encoding a polypeptide of claim
 2. 5. An isolatedpolynucleotide of claim 4 selected from the group consisting of SEQ IDNO:28-54.
 6. A recombinant polynucleotide comprising a promoter sequenceoperably linked to a polynucleotide of claim
 3. 7. A cell transformedwith a recombinant polynucleotide of claim
 6. 8. A transgenic organismcomprising a recombinant polynucleotide of claim
 6. 9. A method forproducing a polypeptide of claim 1, the method comprising: a) culturinga cell under conditions suitable for expression of the polypeptide,wherein said cell is transformed with a recombinant polynucleotide, andsaid recombinant polynucleotide comprises a promoter sequence operablylinked to a polynucleotide encoding the polypeptide of claim 1, and b)recovering the polypeptide so expressed.
 10. An isolated antibody whichspecifically binds to a polypeptide of claim
 1. 11. An isolatedpolynucleotide comprising a polynucleotide sequence selected from thegroup consisting of: a) a polynucleotide sequence selected from thegroup consisting of SEQ ID NO:28-54, b) a naturally occurringpolynucleotide sequence having at least 90% sequence identity to apolynucleotide sequence selected from the group consisting of SEQ IDNO:28-54, c) a polynucleotide sequence complementary to a), d) apolynucleotide sequence complementary to b), and e) an RNA equivalent ofa)-d).
 12. An isolated polynucleotide comprising at least 60 contiguousnucleotides of a polynucleotide of claim
 11. 13. A method for detectinga target polynucleotide in a sample, said target polynucleotide having asequence of a polynucleotide of claim 11, the method comprising: a)hybridizing the sample with a probe comprising at least 20 contiguousnucleotides comprising a sequence complementary to said targetpolynucleotide in the sample, and which probe specifically hybridizes tosaid target polynucleotide, under conditions whereby a hybridizationcomplex is formed between said probe and said target polynucleotide orfragments thereof, and b) detecting the presence or absence of saidhybridization complex, and, optionally, if present, the amount thereof.14. A method of claim 13, wherein the probe comprises at least 60contiguous nucleotides.
 15. A method for detecting a targetpolynucleotide in a sample, said target polynucleotide having a sequenceof a polynucleotide of claim 11, the method comprising: a) amplifyingsaid target polynucleotide or fragment thereof using polymerase chainreaction amplification, and b) detecting the presence or absence of saidamplified target polynucleotide or fragment thereof, and, optionally, ifpresent, the amount thereof.
 16. A composition comprising an effectiveamount of a polypeptide of claim 1 and a pharmaceutically acceptableexcipient.
 17. A composition of claim 16, wherein the polypeptidecomprises an amino acid sequence selected from the group consisting ofSEQ ID NO:1-27.
 18. A method for treating a disease or conditionassociated with decreased expression of functional TRICH, comprisingadministering to a patient in need of such treatment the composition ofclaim
 16. 19. A method for screening a compound for effectiveness as anagonist of a polypeptide of claim 1, the method comprising: a) exposinga sample comprising a polypeptide of claim 1 to a compound, and b)detecting agonist activity in the sample.
 20. A composition comprisingan agonist compound identified by a method of claim 19 and apharmaceutically acceptable excipient.
 21. A method for treating adisease or condition associated with decreased expression of functionalTRICH, comprising administering to a patient in need of such treatment acomposition of claim
 20. 22. A method for screening a compound foreffectiveness as an antagonist of a polypeptide of claim 1, the methodcomprising: a) exposing a sample comprising a polypeptide of claim 1 toa compound, and b) detecting antagonist activity in the sample.
 23. Acomposition comprising an antagonist compound identified by a method ofclaim 22 and a pharmaceutically acceptable excipient.
 24. A method fortreating a disease or condition associated with overexpression offunctional TRICH, comprising administering to a patient in need of suchtreatment a composition of claim
 23. 25. A method of screening for acompound that specifically binds to the polypeptide of claim 1, saidmethod comprising the steps of: a) combining the polypeptide of claim 1with at least one test compound under suitable conditions, and b)detecting binding of the polypeptide of claim 1 to the test compound,thereby identifying a compound that specifically binds to thepolypeptide of claim
 1. 26. A method of screening for a compound thatmodulates the activity of the polypeptide of claim 1, said methodcomprising: a) combining the polypeptide of claim 1 with at least onetest compound under conditions permissive for the activity of thepolypeptide of claim 1, b) assessing the activity of the polypeptide ofclaim 1 in the presence of the test compound, and c) comparing theactivity of the polypeptide of claim 1 in the presence of the testcompound with the activity of the polypeptide of claim 1 in the absenceof the test compound, wherein a change in the activity of thepolypeptide of claim 1 in the presence of the test compound isindicative of a compound that modulates the activity of the polypeptideof claim
 1. 27. A method for screening a compound for effectiveness inaltering expression of a target polynucleotide, wherein said targetpolynucleotide comprises a sequence of claim 5, the method comprising:a) exposing a sample comprising the target polynucleotide to a compound,under conditions suitable for the expression of the targetpolynucleotide, b) detecting altered expression of the targetpolynucleotide, and c) comparing the expression of the targetpolynucleotide in the presence of varying amounts of the compound and inthe absence of the compound.
 28. A method for assessing toxicity of atest compound, said method comprising: a) treating a biological samplecontaining nucleic acids with the test compound; b) hybridizing thenucleic acids of the treated biological sample with a probe comprisingat least 20 contiguous nucleotides of a polynucleotide of claim 11 underconditions whereby a specific hybridization complex is formed betweensaid probe and a target polynucleotide in the biological sample, saidtarget polynucleotide comprising a polynucleotide sequence of apolynucleotide of claim 11 or fragment thereof; c) quantifying theamount of hybridization complex; and d) comparing the amount ofhybridization complex in the treated biological sample with the amountof hybridization complex in an untreated biological sample, wherein adifference in the amount of hybridization complex in the treatedbiological sample is indicative of toxicity of the test compound.
 29. Adiagnostic test for a condition or disease associated with theexpression of TRICH in a biological sample, the method comprising: a)combining the biological sample with an antibody of claim 10, underconditions suitable for the antibody to bind the polypeptide and form anantibody:polypeptide complex, and b) detecting the complex, wherein thepresence of the complex correlates with the presence of the polypeptidein the biological sample.
 30. The antibody of claim 10, wherein theantibody is: a) a chimeric antibody, b) a single chain antibody, c) aFab fragment, d) a F(ab′)₂ fragment, or e) a humanized antibody.
 31. Acomposition comprising an antibody of claim 10 and an acceptableexcipient.
 32. A method of diagnosing a condition or disease associatedwith the expression of TRICH in a subject, comprising administering tosaid subject an effective amount of the composition of claim
 31. 33. Acomposition of claim 31, wherein the antibody is labeled.
 34. A methodof diagnosing a condition or disease associated with the expression ofTRICH in a subject, comprising administering to said subject aneffective amount of the composition of claim
 33. 35. A method ofpreparing a polyclonal antibody with the specificity of the antibody ofclaim 10, the method comprising: a) immunizing an animal with apolypeptide having an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27, or an immunogenic fragment thereof, underconditions to elicit an antibody response, b) isolating antibodies fromsaid animal, and c) screening the isolated antibodies with thepolypeptide, thereby identifying a polyclonal antibody which bindsspecifically to a polypeptide having an amino acid sequence selectedfrom the group consisting of SEQ ID NO:1-27.
 36. An antibody produced bya method of claim
 35. 37. A composition comprising the antibody of claim36 and a suitable carrier.
 38. A method of making a monoclonal antibodywith the specificity of the antibody of claim 10, the method comprising:a) immunizing an animal with a polypeptide having an amino acid sequenceselected from the group consisting of SEQ ID NO: 1-27, or an immunogenicfragment thereof, under conditions to elicit an antibody response, b)isolating antibody producing cells from the animal, c) fusing theantibody producing cells with immortalized cells to form monoclonalantibody-producing hybridoma cells, d) culturing the hybridoma cells,and e) isolating from the culture monoclonal antibody which bindsspecifically to a polypeptide having an amino acid sequence selectedfrom the group consisting of SEQ ID NO:1-27.
 39. A monoclonal antibodyproduced by a method of claim
 38. 40. A composition comprising theantibody of claim 39 and a suitable carrier.
 41. The antibody of claim10, wherein the antibody is produced by screening a Fab expressionlibrary
 42. The antibody of claim 10, wherein the antibody is producedby screening a recombinant immunoglobulin library.
 43. A method ofdetecting a polypeptide having an amino acid sequence selected from thegroup consisting of SEQ ID NO:1-27 in a sample, the method comprising:a) incubating the antibody of claim 10 with a sample under conditions toallow specific binding of the antibody and the polypeptide, and b)detecting specific binding, wherein specific binding indicates thepresence of a polypeptide having an amino acid sequence selected fromthe group consisting of SEQ ID NO:1-27 in the sample.
 44. A method ofpurifying a polypeptide having an amino acid sequence selected from thegroup consisting of SEQ ID NO:1-27 from a sample, the method comprising:a) incubating the antibody of claim 10 with a sample under conditions toallow specific binding of the antibody and the polypeptide, and b)separating the antibody from the sample and obtaining the purifiedpolypeptide having an amino acid sequence selected from the groupconsisting of SEQ ID NO:1-27.
 45. A polypeptide of claim 1, comprisingthe amino acid sequence of SEQ ID NO:1.
 46. A polypeptide of claim 1,comprising the amino acid sequence of SEQ ID NO:2.
 47. A polypeptide ofclaim 1, comprising the amino acid sequence of SEQ ID NO:3.
 48. Apolypeptide of claim 1, comprising the amino acid sequence of SEQ IDNO:4.
 49. A polypeptide of claim 1, comprising the amino acid sequenceof SEQ ID NO:5.
 50. A polypeptide of claim 1, comprising the amino acidsequence of SEQ ID NO:6.
 51. A polypeptide of claim 1, comprising theamino acid sequence of SEQ ID NO:7.
 52. A polypeptide of claim 1,comprising the amino acid sequence of SEQ ID NO:8.
 53. A polypeptide ofclaim 1, comprising the amino acid sequence of SEQ ID NO:9.
 54. Apolypeptide of claim 1, comprising the amino acid sequence of SEQ IDNO:10.
 55. A polypeptide of claim 1, comprising the amino acid sequenceof SEQ ID NO:11.
 56. A polypeptide of claim 1, comprising the amino acidsequence of SEQ ID NO:12.
 57. A polypeptide of claim 1, comprising theamino acid sequence of SEQ ID NO:13.
 58. A polypeptide of claim 1,comprising the amino acid sequence of SEQ ID NO:14.
 59. A polypeptide ofclaim 1, comprising the amino acid sequence of SEQ ID NO:15.
 60. Apolypeptide of claim 1, comprising the amino acid sequence of SEQ IDNO:16.
 61. A polypeptide of claim 1, comprising the amino acid sequenceof SEQ ID NO:17.
 62. A polypeptide of claim 1, comprising the amino acidsequence of SEQ ID NO:18.
 63. A polypeptide of claim
 1. comprising theamino acid sequence of SEQ ID NO:19.
 64. A polypeptide of claim 1,comprising the amino acid sequence of SEQ ID NO:20.
 65. A polypeptide ofclaim 1, comprising the amino acid sequence of SEQ ID NO:21.
 66. Apolypeptide of claim 1, comprising the amino acid sequence of SEQ IDNO:22.
 67. A polypeptide of claim 1, comprising the amino acid sequenceof SEQ ID NO:23.
 68. A polypeptide of claim 1, comprising the amino acidsequence of SEQ ID NO:24. 69 A polypeptide of claim 1, comprising theamino acid sequence of SEQ ID NO:25.
 70. A polypeptide of claim 1,comprising the amino acid sequence of SEQ ID NO:26.
 71. A polypeptide ofclaim 1, comprising the amino acid sequence of SEQ ID NO:27. 72 Apolynucleotide of claim 11, comprising the polynucleotide sequence ofSEQ ID NO:28
 73. A polynucleotide of claim 11, comprising thepolynucleotide sequence of SEQ ID NO:29.
 74. A polynucleotide of claim11, comprising the polynucleotide sequence of SEQ ID NO:30.
 75. Apolynucleotide of claim 11, comprising the polynucleotide sequence ofSEQ ID NO:31.
 76. A polynucleotide of claim 11, comprising thepolynucleotide sequence of SEQ ID NO:32.
 77. A polynucleotide of claim11, comprising the polynucleotide sequence of SEQ ID NO:33.
 78. Apolynucleotide of claim 11, comprising the polynucleotide sequence ofSEQ ID NO:34.
 79. A polynucleotide of claim 11, comprising thepolynucleotide sequence of SEQ ID NO:35
 80. A polynucleotide of claim11, comprising the polynucleotide sequence of SEQ ID NO:36.
 81. Apolynucleotide of claim 11, comprising the polynucleotide sequence ofSEQ ID NO:37.
 82. A polynucleotide of claim 11, comprising thepolynucleotide sequence of SEQ ID NO:38.
 83. A polynucleotide of claim11, comprising the polynucleotide sequence of SEQ ID NO:39.
 84. Apolynucleotide of claim 11, comprising the polynucleotide sequence ofSEQ ID NO:40.
 85. A polynucleotide of claim 11 comprising thepolynucleotide sequence of SEQ ID NO:41.
 86. A polynucleotide of claim11, comprising the polynucleotide sequence of SEQ ID NO:42.
 87. Apolynucleotide of claim 11, comprising the polynucleotide sequence ofSEQ ID NO:43.
 88. A polynucleotide of claim 11, comprising thepolynucleotide sequence of SEQ ID NO:44.
 89. A polynucleotide of claim11, comprising the polynucleotide sequence of SEQ ID NO:45.
 90. Apolynucleotide of claim 11, comprising the polynucleotide sequence ofSEQ ID NO:46.
 91. A polynucleotide of claim 11, comprising thepolynucleotide sequence of SEQ ID NO:47.
 92. A polynucleotide of claim11, comprising the polynucleotide sequence of SEQ ID NO:48.
 93. Apolynucleotide of claim 11, comprising the polynucleotide sequence ofSEQ ID NO:49.
 94. A polynucleotide of claim 11, comprising thepolynucleotide sequence of SEQ ID NO:50.
 95. A polynucleotide of claim11, comprising the polynucleotide sequence of SEQ ID NO:51.
 96. Apolynucleotide of claim 11, comprising the polynucleotide sequence ofSEQ ID NO:52.
 97. A polynucleotide of claim 11, comprising thepolynucleotide sequence of SEQ ID NO:53.
 98. A polynucleotide of claim11, comprising the polynucleotide sequence of SEQ ID NO:54.
 99. A methodof claim 9, wherein the polypeptide has the sequence of SEQ ID NO:1.100. A method of claim 9, wherein the polypeptide has the sequence ofSEQ ID NO:2.
 101. A method of claim 9, wherein the polypeptide has thesequence of SEQ ID NO:3.
 102. A method of claim 9, wherein thepolypeptide has the sequence of SEQ ID NO:4.
 103. A method of claim 9,wherein the polypeptide has the sequence of SEQ ID NO:5.
 104. A methodof claim 9, wherein the polypeptide has the sequence of SEQ ID NO:6.105. A method of claim 9, wherein the polypeptide has the sequence ofSEQ ID NO:7.
 106. A method of claim 9, wherein the polypeptide has thesequence of SEQ ID NO:8.
 107. A method of claim 9, wherein thepolypeptide has the sequence of SEQ ID NO:9.
 108. A method of claim 9,wherein the polypeptide has the sequence of SEQ ID NO:10.
 109. A methodof claim 9, wherein the polypeptide has the sequence of SEQ ID NO:11.110. A method of claim 9, wherein the polypeptide has the sequence ofSEQ ID NO:12.
 111. A method of claim 9, wherein the polypeptide has thesequence of SEQ ID NO:13.
 112. A method of claim 9, wherein thepolypeptide has the sequence of SEQ ID NO:14.
 113. A method of claim 9,wherein the polypeptide has the sequence of SEQ ID NO:15.
 114. A methodof claim 9, wherein the polypeptide has the sequence of SEQ ID NO:16.115. A method of claim 9, wherein the polypeptide has the sequence ofSEQ ID NO:17.
 116. A method of claim 9, wherein the polypeptide has thesequence of SEQ ID NO:18.
 117. A method of claim 9, wherein thepolypeptide has the sequence of SEQ ID NO:19.
 118. A method of claim 9,wherein the polypeptide has the sequence of SEQ ID NO:20.
 119. A methodof claim 9, wherein the polypeptide has the sequence of SEQ ID NO:21.120. A method of claim 9, wherein the polypeptide has the sequence ofSEQ ID NO:22.
 121. A method of claim 9, wherein the polypeptide has thesequence of SEQ ID NO:23.
 122. A method of claim 9, wherein thepolypeptide has the sequence of SEQ ID NO:24.
 123. A method of claim 9,wherein the polypeptide has the sequence of SEQ ID NO:25.
 124. A methodof claim 9, wherein the polypeptide has the sequence of SEQ ID NO:26.125. A method of claim 9, wherein the polypeptide has the sequence ofSEQ ID NO:27.
 126. A microarray wherein at least one element of themicroarray is a polynucleotide of claim
 12. 127. A method for generatinga transcript image of a sample which contains polynucleotides, themethod comprising the steps of: a) labeling the polynucleotides of thesample, b) contacting the elements of the microarray of claim 126 withthe labeled polynucleotides of the sample under conditions suitable forthe formation of a hybridization complex, and c) quantifying theexpression of the polynucleotides in the sample.
 128. An arraycomprising different nucleotide molecules affixed in distinct physicallocations on a solid substrate, wherein at least one of said nucleotidemolecules comprises a first oligonucleotide or polynucleotide sequencespecifically hybridizable with at least 30 contiguous nucleotides of atarget polynucleotide, said target polynucleotide having a sequence ofclaim
 11. 129. An array of claim 128, wherein said first oligonucleotideor polynucleotide sequence is completely complementary to at least 30contiguous nucleotides of said target polynucleotide.
 130. An array ofclaim 128, wherein said first oligonucleotide or polynucleotide sequenceis completely complementary to at least 60 contiguous nucleotides ofsaid target polynucleotide.
 131. An array of claim 128, which is amicroarray.
 132. An array of claim
 128. further comprising said targetpolynucleotide hybridized to said first oligonucleotide orpolynucleotide.
 133. An array of claim 128, wherein a linker joins atleast one of said nucleotide molecules to said solid substrate.
 134. Anarray of claim 128, wherein each distinct physical location on thesubstrate contains multiple nucleotide molecules having the samesequence, and each distinct physical location on the substrate containsnucleotide molecules having a sequence which differs from the sequenceof nucleotide molecules at another physical location on the substrate.